TREE PROTECTIONNEWS

NewsLetteV" of tne TV"ee 'FYotectLoV'vco-o-pen/ltLve PV"og n::JVvtVvte

VlV'vLveV"sLtuof PV"etoV"LCl

MAY200SNO. 5

CONTENTS

The Director reporting

Field Services Manager receives covetedaward

L'oreal-UNESCO fellowship for young~m~ 3

Annual TPCP meeting March 2003 4

Executive summary ofTPCP activities 4

New Forest Molecular genetics programme inFABI 5

Fungi associated with Conifer-infesting BarkBeetles in Norway and South Africa 6

Welcome to the TPCP/FABI

International visitors to TPCP

TPCP Diagnostic Clinic 2003

Expanding our wood decay studies to utilitypoles 10

The Research Team of the TPCP

Contacting the TPCP: Your 2-Stop-Shop 11

Bilateral agreement between the Belgium andSouth African governments 11

The Bluestain puzzle 12

Sirex: The road ahead 12

International Congress of Plant Pathology(ICPP) 13

Abstracts from the recent InternationalCongress for Plant Pathology

FABI becoming Fabylon

Graduations

10

14

15

16

2

The status of plantation health appears to be changing and presenting us withnew and unexpected challenges. This worrying change of environment beganto emerge in recent years with growing damage due to the pitch cankerpathogen, Fusarium circinatum. During the course of the last year, the Sirexwood wasp Sirex noctilio with its associated fungal symbiont, Amylostereumareolatum, began to appear in pine plantations far north of the previouslyrecognised northern limits. Dealing with these two problems will present us

The Directorreporting .As I write this note, I am flying down to Richards Bay to meet withTree Protection Co-operative Programme (TPCP) Field Services Manager,Jolanda Roux and overseas guests that attended our annual meeting thislast week. What lies ahead is a few days of lectures to forestry colleagues andfield days to inspect trials in the Zululand area. As one might expect from thistime of the year, it is likely to be hot and muggy in the field, but on the morepositive side, the disease and pest problems will be interesting and exciting toconsider. In addition, having the opportunity to hear the views of overseaspathology and entomology experts regarding our various projects is a specialprivilege, and always much appreciated.

6

With the TPCP meetings now over, the research and extension team can getback to normal activities. I believe that all will agree that this years meetingwas again a great success. As important as the meeting is, I have to admit thatit is a relief to have it behind us for another year. This event requires a hugeamount of work to organise, and some elements of the planning occur up to ayear in advance. Dates are now set for our 2004 meeting (April 20, 21 and 22)and venues are already booked. Some three months before the meeting occurs,the team enters into a period of intense activity and this does not change untilthe meeting is over. So while we all feel elated at the outcome of the meetings,I would be untruthful to say that we are not somewhat pleased that it is over fornow.

7

8

At this time, I cannot help but reflect on the overall outcome of the TPCPmeeting. I will not comment on the regular events - the presentations by teamresearches etc- but rather on some of the special features. Here, presentationsby Carl Seele (NCT), Andrew Morris (Sappi), Bruce Hulett (Mondi), CarolRolando (ICFR) and Glen Mitchell (Fusarium Working Group) deserve specialmention. All five presentations were outstanding and brought specialperspective to the links between the TPCP team and our forest industrypartners. In addition, we enjoyed fantastic presentations from Dr. Giles Hardyform Murdoch University in Perth, Dr. Kier Klepzig from the USDA ForestService research station in Pineville, Louisiana and Mrs. Nora Telechea ofDirection Forestal, Montevideo, Uruguay. Their perspectives on Eucalyptus'and Pine pests and diseases were important and valuable.

with huge challenges in coming years. For now, a verypositive sign is that all players in the industry are committedto fighting the battle. Here the Fusarium Working Group hasbeen and will increasingly be extremely important. Likewise,·during the past few weeks, an all-industry Sirex Action Plan(SAP) steering committee and a SAP technical committeehave been constituted. Together with the TPCP, thesecommittees will forge ahead with the management of theSirex threat.

There can be little doubt that new pests and diseases willchallenge forestry in South Africa in the future. Sirex and thepitch canker fungus are just two examples of what we mightexpect in years to come. This should not be interpreted as agloomy and pessimistic perspective, but rather as a realisticchallenge that will need to be met. From this standpoint, thepitch canker fungus problem and the Sirex wood wasp threatpresent us with positive as well as negative elements. Whilethey are certainly serious, they enable us to learn to deal withother new and serious incursions that we will need to dealwith in the future.

Before ending this introductory note, I must take thisopportunity to thank all of you that attended this year's annualTPCP meeting. My commiserations to those people that hadplanned to attend and were prevented from doing so due tofires in plantations. This event is a special feature in theSouth African Forest Industry calendar and brings manypositive products to our industry. Among these are open linesof communication between researchers and field foresters aswell as between staff of the many groups that make upForestry in this country today. Such communication isessential in our quest towards the goal of "KEEPING TREESHEALTHY"

Mike Wingfield.**************************************************

FIELD SERVICESMANAGER RECEIVES

COVETED AWARDDr. :rolanda Roux, well known to most forestersand forest managers in South Africa has receivedone of the coveted "Exceptional YoungResearcher Awards" from the University ofPretoria. About four of these awards are made toyoung researchers each year and they recogniseexceptional research achievements. For Jolanda,these achievements are many and varied. She hasparticularly been a pioneer in elucidating diseasesof plantation trees in various countries of Africa,including South Africa. She has also conductedgroundbreaking research on diseases of black wattleand especially on the serious Ceratocystis wiltpathogens of wattle and eucalypts. As a youngresearcher, she has also played a key role inadvising post-graduate students and, in this regard,her first M.Sc. and Ph.D. students have justcompleted their studies. The following citation waspresented with the award that comes with acertificate and monetary incentive:

"Dr. J. Roux conducts research on diseases of trees,particularly those caused by fungi. Much of her

research has, been on thediseases affecting commercialplantation forestry in SouthAfrica, but she has also doneresearch into the diseasesaffecting forestry in Africancountries such as Uganda andEthiopia. Her research includesthe identification of pathogensusing morphological andmolecular techniques. She alsomakes use of phylogenetic andpopulation studies to betterunderstand the pathogens andassists in the selection ofdisease-tolerant plantingmaterial for South Africanforestry companies."

DO NOT .. I REPEA T ..DO NOT! cut the tree whIle I'm up here collecting samples!!(The manager of field services up a tree)

2

Dr. Karin Jacobs is the second SouthAfrican woman to become the recipient ofthe annual International L'oreal UNESCOfellowship for young women in science.L'oreol. in partnership with UNESCO, introduced theaward programme in 1998 to recognize thecontributions of exceptional female scientists and toencourage promising young women scientists fromaround the world. Currently, five awards are given towomen representing the five geographical regions(Africa, Arab States, Europe and North America,Asia-Pacifica and Latin America) in recognition oftheir contribution to their scientific field. Fifteenpost-doctoral fellowships are awarded to promisingyoungwomenat the start of their careers. NationalCommissions of each country can nominate twocandidates for the fellowships and an internationaljury makes the final selection from thesenominations. The fellowships were presented to therecipients at a glamorous award ceremony at theUNESCO headquarters in Paris on the 27th ofFebruary 2003.

Karin is currently working as a post-doctoral fellow inthe TPCPprogramme under the supervision of Prof.Brenda Wingfield. She is no stranger to theprogramme and completed her PhD with Prof. MikeWingfield before spending two years withAgriculture Agrifood Canadain the laboratory of Dr.Keith Seifert. Her main research interests at themomentare to compilea multigene phylogenyfor thegenusLeptographium and to revise the monographofLeptographium, published in 2000. These fungi arepart of the group that causesblue-stain on lumber.

The fellowship will commencein October 2003 andwill be aimed at the development of a rapididentification method for fungal pathogensbased onmicro-array technology. This method is basedon the

exploitation of short pieces of DNA(oligonucleotides) uniqueto a certain species, fungalpathogens in this case. Single strand copies ofthese unique oligonucleotides are attached to amicroscope slide with the aid of a micro-arrayspotter. A hybridisation reaction with a samplecontaining unknownfungi will result in the formationof double strands where the sample DNA iscomplementary to the oligonucleotides on the slide.This will result in a colour reaction, indicating apositive identification. If the sampleDNA containsno DNA that is complimentary to the target DNA,there will be no colour reaction. The advantage isthat the technique is very rapid and accurate. Afurther advantage is that one can test a singlesamplefor 500-600 species at the sametime. Theproposed project will be done in collaboration withDr. C.A. Levesqueand Dr. Seifert from AgricultureAgrifood Canada,who is already in the process ofdeveloping"microchips" for Canadianagriculture andforestry.

3

ANNUALTPCPMEETING MARCH2003On the 11th and 1th of March 2003 the TPCP had its annualmeeting at the University of Pretoria. Despite a few peoplemissing the meeting at the last moment due to plantation fires,we once again had excellent attendance at the meeting. Forthose who were not lucky enough to attend we have decided topublish at least the executive summary of some of the activitiesof the TPCP in this issue of our newsletter.

EXECUTIVE SUMMARYOF TPCP ACTIVITIES

• This report marks the end of twelve years of formalexistence of the TPCP. During this time, the Programme hasgrown considerably, both in size and stature. It hascontinuously changed to meet the needs of the South Africanforestry industry and to ensure that diseases and more recently,insect pests do not impinge on the sustainability or reduce theproductivity of plantation forestry in South Africa. It is nowwidely accepted internationally that the TPCP is one of thestrongest and most active unified programmes dealing withplantation pests and diseases.

• Diseases and pests pose one of the greatest threats toplantation forestry in South Africa and elsewhere. Theprediction that losses due to these perturbations will increasehas been highlighted in recent years by the appearance of anuncomfortably large number of pests and pathogens. There areclear indications that this trend will continue and great effortwill be needed to ensure that losses are minimal.

• During the past year, the TPCP has continued tomaintain an active programme of disease and pest monitoring.This has been achieved through field surveys, participation inmonitoring activities of members, training of field forestersthrough field days and extension and through maintaining anactive pest and disease diagnostic clinic that provides a freeadvisory service to members. During the past year, the teamspent 635 person days in member plantations and this high levelof activity in the field is more or less equal to that in past years.

• Being housed in a University implies that the TPCPhas a primary responsibility to train post graduate students inthe various disciplines related to tree protection. From aUniversity standpoint, the group has experienced an exceptionalyear with a record number of students completing M.Sc. andPh.D. degrees and an outstanding record of publishing highquality science in internationally reputable journals. Bymaintaining this exceptional record, the group is easily able toretain the strongest possible support from the University ofPretoria.

• During the past year, the University reaffirmed itssupport to the group by approving a R20 million grant toexpand the facilities used by the TPCP. Thus, during thecoming year, the "downstairs" FABI facilities will beextended by more than double its current size. The building

project is well advanced and should be completed by mid2003. This will include additional laboratories, facilities forworking with genetically modified plants and the second onlyquarantine level greenhouses in South Africa. These newfacilities will have a great impact on the work that theProgramme is able to undertake.

• Many pest and disease problems were dealt withduring the last year and details of the most important of theseare provided in the extended report to members. However twoparticular problems caused the highest level of concern andthese have required the greatest effort from the TPCP team.More specifically they include the damage due to the pitchcanker fungus, Fusarium circinatum and the Sirex wood wasp,Sirex noctilio. Special programmes have been initiated to dealwith these agents that seriously threaten forestry in SouthAfrica.

• New diseases have been discovered for the first timein South Africa and many well-established diseases have beensubjected to intensive study. Of the new pathogens discovered,we are most concerned about the appearance of the serious wiltpathogen Ceratocystis fimbriata. This fungus has led to thedeaths of large numbers of Eucalyptus trees elsewhere in theworld, and a special effort will need to be made to deal withthis newly discovered tree pathogen in South Africa.

• During 2002, the TPCP has continued with its effortsto integrate its pathology and entomology activities. This hasnot been difficult to achieve because the requirements of thedisciplines, especially in forestry, are so similar. Projects arealready closely integrated and will continue to be more so inthe coming years.

• The TPCP has continued to maintain a very activeprogramme of international collaboration. This is mostimportant, especially in the field of tree protection where it isnecessary to maintain a clear view of problems beingexperienced elsewhere in the world. Pests and pathogens aremoving actively and a first hand understanding and awarenessof key problems must facilitate early discovery, risk abatementand reduced losses. Amongst the formal collaborations that weare proud to have continued are those partially funded by theAustralian government to evaluate the risks of Eucalyptus rust,and bilateral agreements between the South African andChinese Governments, the South African and NorwegianGovernments and the South Africa and Belgium Governments.

• Members of the TPCP team have excelled in thenational and international arenas, receiving a wide range ofawards and invitations. We are particularly pleased with theseawards and invitations in the case of students, many of whomhave gained excellent funding opportunities, together withexceptional new experiences.

• As in the past, the successes of the TPCP are linked toa high level of collaboration and team effort on the part of agreat number of people. The research group and support staffof about 50 represents an outstanding team of highlycommitted and passionate people. The group gains tremendoussupport from University colleagues and especially forestmanagers and foresters throughout South Africa. We take thisopportunity to thank members of the TPCP and their staff forvalued and superb support. It is only though this support andcollaboration that we are able to achieve our goal of"KEEPING TREES HEALTHY".

4

NEW FOREST MOLECULARGENETICS PROGRAMME INFABI STARTS WITH HELPOF SAPPI FORESTS

Two automated DNA analysers were recently installed at theUniversity of Pretoria. These instruments were purchasedfrom LI-COR Biosciences (Lincoln Nebraska) with a donationof R150 000 from Sappi Forestry and financial contributionsfrom the University of Pretoria, Departments of Genetics,Microbiology and Plant Pathology and the Tree Pathology Co-operative Programme. The LI-COR instruments will be usedfor high-throughput, automated marker analysis of thegenomes of forest trees, cereal crops, banana, fungalpathogens and bacteria. This capacity will form an importantpart of the new Forest Molecular Genetics programme in

Zander Myburg (right) with the first two MScstudents (Martin Ranik and Minique de Castro) in theForest Molecular Genetics programme.

FABI. The programme will be directed by Dr. Zander Myburgwho joined the University of Pretoria at the end of 2001 aftercompleting a PhD degree programme in Forest Biotechnologyat North Carolina State University.

The Forest Molecular Genetics programme will have threemain focus areas. The first focus will be the development ofgeneral molecular tools for forest tree breeding. This includesthe use of molecular markers for fingerprinting of trees andclones, paternity analysis, pollen contamination studies, etc.The second focus will be the development of trait-linkedmolecular markers for marker-assisted improvement of woodand fibre properties, growth and volume, vegetativepropagation and disease resistance. Association geneticsapproaches will be used to identify these genes in treebreeding populations. Thirdly, the programme will focus on

The two LI-COR instruments installed at the Department of Genetics at UP

the identification and functional analysis of genes that areinvolved in wood formation in forest trees. The sequence ofthe poplar genome will be completed within the next year andthis important development in Forest Genomics is expected tolead to a rapid expansion of our knowledge about genes thatregulate wood and fibre development, as well as importantbiological processes such as resistance to forest pathogens.The Forest Molecular Genetics programme at FABI will usecutting-edge technologies such as microarray analysis andAFLP transcript profiling to identify genes involved in thesebiological processes. The function of these genes will bestudied in Eucalyptus trees and in the model plant Arabidopsisthaliana. This tiny weed, of which the complete genomesequence is known, can be induced to form wood and cantherefore be used as a model system for wood and fibredevelopment. Genes characterized in this model willeventually be used to improve plantation forests using marker-assisted breeding or direct genetic engineering.

Zander Myburg collecting differentiating xylem (wood formingtissue) from a Eucalyptus tree

5

For more information on Forest MolecularGenetics visit:http://www.up.ac.zaJacademic/fab~/eucgenomics/Eucalyptus.htm

FUNGI ASSOCIATED WITHCONIFER-INFESTING BARKBEETLES IN NORWAY AND

SOUTH AFRICAA South African-Norwegian Research

Cooperation

Bark beetles, particularly in the genera /ps and Hy/astesare common in Europe, including Norway, and representone of the greatest threats to conifer plantations, as theycarry a guild of fungi that contribute to tree death. Theseinsects are also important to South Africa, becauseseveral species have been accidentally introduced intothe country. Much work has been done on bark beetle-associated fungi in Norway, but little is known regardingthese organisms in South Africa. A new 1.5 million randproject funded by the South African - NorwegianProgramme for Research Cooperation aims to improveon this situation by exchanging knowledge andresearchers between the two countries. The Norwegianpartner, the Norwegian Forest Research Institute(Skogforsk), has a long history of forestry research andis particularly well-known for the work that has beendone in the areas of classical forest pathology and forestentomology. FABI has a strong history of research ondiseases of plants and trees, with particular emphasis onmolecular studies of tree pathogens.

The main objective of this collaborative project is tocharacterise the fungal pathogens associated with anumber of important bark beetles in Norway and relatedspecies that have been accidentally introduced intoSouth Africa. The research will contribute to anunderstanding of bark beetle biology, the identification offungi associated with bark beetles, as well as the impactof bark beetles and their fungal associates on foresttrees. Specific objectives are:

1 . To identify fungi associated with different species ofbark beetles in South Africa and Norway, using bothconventional and molecular techniques such as DNAsequencing. Some of the fungi have not been characterisedand poor identifications are already impacting negatively ontimber trade, which is important to both Norway and SouthAfrica.

WELCOME TOTPCP/FABI

Mohamined Abdo has started his Masters degreeon the development of micro satellite markers forFusarium circinatum. Mohammed is originallyfrom Egypt where he obtained his B.Sc. degree.

2. To develop molecular markers for species ofdominant fungi to enable comparisons of populations.Through undertaking population biology studies, we hope todiscover the origins of fungi associated with introduced barkbeetles in South Africa. This will aid in improving quarantinemeasures both in South Africa and Norway. Patterns of spreadof the fungi and their insect vectors may also be elucidated.

3. To test the pathogenicity of selected fungi in SouthAfrica using techniques developed at Skogforsk. Very little isknown about the pathogenicity of fungi associated withinvasive conifer bark beetles in South Africa. The pathogen-icity of blue-stain fungi is studied using artificial pointinoculation of fungal mycelium into host trees. Trees areeither inoculated at low densities, or mass inoculated with upto 300 inoculations per tree. With a low number ofinoculations the fungus is always confined by the defenceresponses of the tree. The mass-inoculation technique, whichhas been developed at Skogforsk, simulates the depletion ofhost defenses that takes place during a bark beetle mass-attack, and mature, healthy trees can be killed using thistechnique.

The two partner institutions have mutual andcomplimentary interest in this research domain. TheNorwegian partners bring to the project the long-termexperience and knowledge of bark beetle biology andthe fungi associated with these important pests. Closeinteraction between South African students and researchleaders from Norway will be achieved by studentsworking in Norway and the Norwegian partners workingin South Africa together with these students. During thecourse of the 3-year project (running from 01/Aug/02 to31/JuIl05) students and staff will be exchanged betweenFABI and Skogforsk for a total duration of 29 months.Most of this time (23 month) will be allocated to SouthAfrican PhD students and Post does working in Norway.Through this mentorship students will come back toSouth Africa with experience particularly in forestentomology. This is a seriously neglected subject inSouth Africa and is a serious problem that needs urgentattention. We envisage that this project will contributesignificantly to increasing forest entomology knowledgeand capacity in South Africa.

He, however, came to South Africa and recentlycompleted his B.Sc. honours degree inMicrobiology and the University of Pretoria.

Izette Greyling will do her Masters degree in theTPCP and will be working on the Pantoea spp.associated with Coniothyrium canker onEucalyptus. She is a graduate of the University ofPretoria.

6

INTERNA TIONALVISITORS TO

TPCPSince our previous issue of Tree Protection News,the TPCP has hosted many international visitors.These visitors provide a valuable opportunity forour students and staff to increase their knowledgeof the international disease situation. Many ofthese visits have also resulted in valuableinternational collaboration with regards to researchprojects and the exchangeof students. These visitswould not be possible without the help of ourforestry colleagues. A special word of thanks goestoall who assisted us in entertaining these visitors andin showing them all the various aspects of ourplantation health research. Thank you also for helpwith accommodation,mealsandyour precious time.

Top: Dr. PiaBarklund (right) and Leon Labuschagne(left) looking at Coniothyrium stem canker inZululand

Our visitors were:

Dr. Pia Barklund from the World AgroforestryCentre crCRAF) in Nairobi, Kenyavisited the TPCPinOctober 2002. She has spent a number of yearsworking in Africa and before here appointment inNairobi, she worked at the Wondo Genet ForestryCollegein Ethiopia for several years.

Dr. Giles Hardy from Murdoch University in Perth,Western Australia is well-known for his work onPhytophthora cinnamomi on E marginata. Morerecently he has been actively involved in researchinto the diseases affecting the recently establishedplantations of E globulus in Western Australia.

Bottom: Mike Wingfield and international visitorsin Cryphonectria inoculation trial F.I t.r: Dr. KierKlebzig, Mike Wingfield and Dr. G!les Hardy.,(Front) Mrs. Nora Telechea

Dr. Kier Klebzig is from the USDA Forest Serviceresearch station in Pineville, Louisianaand has bothentomologicaland forest pathology training. He hasspent the last few years working especially on themanagementand study of bark beetles infesting pinetrees in the U.S.A.

Mrs. Nora Telechea from Direction Forestal inMontevideo, Uruguay is a forest pathologist, alsowith a lot of experience in forest entomology. Shehas been very active especially in the biologicalcontrol of several of Uruguay'sforestry pests.

7

I I

I~IThe structure of the Clinic was adjusted at the beginning of 2003 in order to reflect theactivities of the team (Fig. 1). It now has a diagnostic and research component. The lattercomponent focuses on research samples, specifically pine, received from the TPCP members aswell as samples from the Pine Fusarium Working Group.

For the period January to 20 March, the clinic has received 79 diagnostic and 267 researchsamples (Fig. 2). Of the diagnostic samples, an equal number of pine and eucalypt sampleswere received (Fig. 3). The "other" samples refer to those of bark media, seeds, water etc.

Figure 1Structure of TPCP Diagnostic Clinic

Entomology:Brett Hurley

• Diagnostic Services

ITeam:Lorenzo LombardElsie de MeyerIzette Greyl ingGrace Nakabonge

1__T_PC_P_20_0_3 _I

~usariulnResearch

Fusarium•••• IDs:

Riana Jacobs

8

Technician:Lydia Twala

Figure 2Number of diagnostic and research samples received (Jan to Mar 2003)

No. ofSamples

16014012010080604020

O~----I.....-...IoIL

Jan Feb MarMonths of the Year

Figure 3NUMBER OF SAMPLES RECEIVED FROMDIFFERENT HOSTS (Jan to Mar 2003)

9

o DiagnosticoResearch

OPineoEucalypts• Wattleo Other

Eskom currently has an estimated 2.2 million wooden utilitypoles in service within it's entire distribution network. Theaverage expected life of a utility pole is between 18-35 yearsdepending on the wood species being used and the exposurearea. Figures from 1998 indicate that the failure rate hasincreased to 5% over a four-year period (i.e. 46 875 woodenpoles per annum). The average cost of replacing a pole isapproximately R250 per l l m pole, excluding the installationcosts, resulting in an additional cost to the company of R12million per annum.

Various factors have been identified as contributing to theearly failure of utility poles:• Lack of quality control procedures on the supply side asthe demand exceeds supply,• Insufficient hardening and drying of the pole prior toinstallation, and• Leaching of chemical preservatives from poles.

Eskom recently approached FABI with arequest to conduct a survey to identifythe causal agents of decay on poles indifferent geographical and climaticregions in South Africa. The aim of theproject, which started in January thisyear, is to make a proper diagnosis ofwhich fungal species are involved in thedecay of poles.

These factors contribute to increased fungal and termite attackon the poles as well as twisting and cracking of poles.Conventional treatment of these poles included chemicaltreatments with fungicides and preservatives such as creosote.At present, these methods seem to be only partially effective.Most of these methods are, furthermore, damaging to theenvironment. Alternative methods, therefore, need to beinvestigated to increase the life span of these utility poles.

Decay of poles is primarily caused by fungi attacking thewood, which leads to a reduction in strength and eventualdisintegration of the wood. Many species of fungi can beinvolved in decay. The fungal species vary between differenttypes of wood, wood treated with different chemicals, andalso in different geographical areas.

In South Africa, utility poles are produced primarily fromeucalypt, pine and wattle trees. A considerable amount ofresearch haS been conducted on the production, strength,

specifications and treatment of poles inSouth Africa during the past 40 years.However, no published data is availableon the fungal species responsible fordecay of poles in South Africa. Most ofthe treatments were considered to beeffective against a broad spectrum ofspecies, but with no knowledge about thecausal agents, it is possible that some ofthese methods might not have beeneffective against all the organismsinvolved.

Elsie checking on a telegraph pole

++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++THE RESEARCH TEAM OF THE

TREE PATHOLOGY CO-OPERATIVE PROGRAMME

The research team of the Tree Protection Co-operative Programme is varied. It includes full time staff of the University ofPretoria: Prof MJ Wingfield, Director and Mondi Professor, Prof B Wingfield, Prof TA Coutinho, Or J Roux, and Or PGovender, Rosemarie Visser, Sonja de Beer, Elizabeth Attinger, Eva Muller, Brett Hurley, Hardus Hatting, Helen Doman,Valentine Nkosi, and Martie van Zyl, colleagues and students attached to other organizations such as the ICFR, technicalassistants funded by the University or through membership fees and post graduate students who are mainly funded bythe NRF. Staff from various Departments in the University obviously provide advice and support where this is required.

++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++10

@wi1rf1@OOOO[f) &lJ@ 7JfP@JJ~C&Lf@~[l{j@{]J

aa

For all members' pest and disease problems please make use of thefollowing two people to assist you.

FIELD ENQUIRIESAND EXTENSION

Dr. Jo/anda RouxEmai/: jo/anda.roux@fabi.up.ac.zaTe/: 082 372 8350/012 420 3938Fax: 012 420 3960

PEST AND DISEASEDIAGNOSTIC CLINIC

Dr. Teresa CoutinhoEmai/: teresa.coutinho@fabi.up.ac.zaTe/: 012 420 3934/8/9Fax: 012 420 3960

Bilateral Agreementbetween the Belgium and

South AfricanGovernments

trees in South Africa. The disease is widespread in thiscountry, occurring on a number of different species, hybridsand clones. Trees younger than 3 years are particularlysusceptible. Repeated infections lead to multi-stemmed trees,and undesirable silvicultural trait.

Specific aims of the collaborative research project are to:

• Develop a rapid, identification technique for P.ananatis

• Identify and characterise isolates of P. ananatisinfecting hosts from other countries, includingeucalypts

• Determine the genetic relatedness between P.ananatis isolates from different hosts

• Identify and characterise the two species of Pantoeaassociated with Coniothyrium canker of eucalypts

• Investigate the synergism that exists between the twoPantoea species and Coniothyrium zuluense.

The TPCP under leadership of Prof. Teresa Coutinho appliedand were recently awarded a two-year grant from an initiativebetween the Flemish Community in Belgium and the SouthAfrica Government. The Agreement funds scientific andtechnological research of mutual interest to both partners. Theresearch collaboration is between the Universities of Gent andPretoria. The research focuses on Pantoea ananatis, thecausal agent of bacterial blight and dieback of young eucalypt

11

For the past sixyears a teamfrom FABI hasbeen researchingthe sapstainproblem on pinelogs exported from South Africa for veneer production. Theselogs are infected with staining fungi and on arrival in southeast Asia the sapwood are stained severely, causing extensivelosses to the companies involved.

Initially, a survey was conducted over a two-year period todetermine the causal agents, the seasonal occurrence of stain,and to evaluate currently applied treatments of the logs. Theresults of this survey showed that logs are stained during allseasons, but that it develops slower during the winter months.Furthermore, we found that although the chemical treatmentinitially slows down stain development, levels of stain werethe same in treated and untreated logs after 4 weeks insummer and 8 weeks in winter.

Most importantly, we identified a number of fungal speciescausing the stain. These included several bark-beetleassociated ophiostomatoid fungi, .which are known to causebluestain all over the world on almost all kinds of wood. Alsoincluded was Diplodia pinea, a well-known endophyte (i.e.living inside the healthy tree) and stress related pathogen ofpines in South Africa. However, to our surprise, we foundthat Lasiodiplodia theobromae was responsible for thestaining of more than 80 % of logs surveyed. This fungus hasbeen recorded previously as a pathogen from many differentspecies of plants. It has also been recorded as the agent ofsapstain on several tropical hardwoods. What is interesting, is

First recorded in South Africa at Cape Town in 1994, thewood wasp Sirex noctilio is rapidly spreading through SouthAfrica's pine forests. Last year Sirex was detected as far eastas Weza, just within KwaZulu-Natal, and it is expected thatSirex will be detected in areas beyond this during the intensivemonitoring programme that began this year. The spread ofSirex is coupled with the absence of its biological controlagents, which have failed to establish and spread with theirhost. The result of the unhindered spread of Sirex is alreadyevident in certain plantations, where some compartments have30% and greater loss due to Sirex. If no action is taken, Sirexassociated mortality can far exceed this and has the potentialto cripple our pine industry.

In response to the serious threat posed by Sirex, FSA (ForestrySouth Africa) has provided funds to the TPCP for theestablishment of a Sirex Control Programme, including theemployment of two full-time staff. These staff, namely MrBrett Hurley and Mr Hardus Hatting, will be responsible forthe establishment and management of the Sirex ControlProgramme. The active involvement and co-operation of theentire forest industry in this programme has recently beensecured. The strategy proposed by the TPCP to form the basisof the Sirex Control Programme was discussed and agreedupon with the industry.

The management strategy consists of six major components.Monitoring is the first component and involves an intensive

that is has never beenrecorded as the agentof sapstain of pinesfrom countries suchas Chile and New

Zealand, whichcompetes with South Africa for the same markets in SouthEast Asia.

Following the survey, 16 antisapstain compounds available onthe South African market were screened in laboratory testsagainst the fungi involved. Based on performance, 8 of thesewere selected for trials in the Richards Bay harbour on 1.5 mlogs. The results showed that the treated logs all had the sameamount of stain as the water treated control logs. The trialswere repeated on 3 m logs with the same disappointingresults. One of the reasons for the failure of the chemicals inlog trials is due to the fact that Lasiodiplodia theobromaegrows much faster than most of the other staining fungi, andfailure to treat logs within hours after felling results in stainpenetrating deeper than the chemicals before it has beenapplied. The chemicals were, furthermore, developed againstfungi penetrating the wood from the cut and debarkedsurfaces. Sapstain fungi reported from the NorthernHemisphere and New Zealand infect logs in this manner. Thewater-based chemicals themselves, however, penetrate thelogs at most 1 cm deep, meaning any fungus already inside thesapwood further away than 1 cm from the surface, is notaffected by the chemical. The possibility of endophyticinfections might also contribute to the difficulty to control thestain. The efforts by FABIANS to understand the biology andinfection strategy of the fungus, which should assist us indeveloping effective control measures, continue.

Sirex:The road

aheadapproach to identify the distribution and infestation levels ofSirex within South Africa. Biological control is the maincomponent of the strategy. This involves the re-introductionof biological control agents in Sirex infested areas and theassessment of their efficiency. The primary biological controlagent has the potential to achieve above 90% parasitism ifeffectively introduced. Sirex awareness aimed at forestrystaff, timber buyers and the general public is needed topromote the basic awareness of Sirex and its situation in SouthAfrica. Quarantine, silviculture and research are the otherthree components of the strategy. Research includes assessingthe biological control agents, obtaining a better understandingof Sirex and our situation in South Africa and investigatingalternative control measures. The strategy will commencefrom this year onwards.

12

A Sirex Programme Steering Committee (SPSC) and SirexProgramme Technical Committee (SPTC) have beenestablished to manage South Africa's Sirex Action Plan. TheSirex Programme Steering Committee initially chaired byProf. Colin Dyer of the ICFR, will oversee the programme.The Sirex Programme Technical Committee will be ASresponsible for the implementation of the strategy and report NEW SFL Hto the Sirex Steering Committee. Both committees include __ --members of industry and government and staff of the TPCP. MARCH 2003 _ SIREXThese committees will work towards the success of the SirexControl Programme in managing the population of Sirex atlevels below concern for the forest industry.

For more information contact Brett Hurleybrett.hurley@fabi.up.ac.za

Cell: 0824508941

FOUND IN HOWICK

FABIITPCP Group attending the ICPP Congress in Christchurch, New ZealandFebruary 2003

The 8th International Congress of PlantPathology (ICPP) was held in Christchurch,otherwise known as the garden city of the world,New Zealand from the 2nd to the 7th of February2003. The theme of this year's congress was "Solvingproblems in the real world". A total of 1300 delegatesattended the congress and they represented 74 countries. Atotal of 1200 poster presentations were exhibited and therewere invited oral presentations on various aspects of plantpathology. The Forestry and Agricultural BiotechnologyInstitute (FABI) and the Tree Pathology Co-operativeProgramme (TPCP) were well represented with a group offourteen postgraduate students and staff in attendance.

There were five keynote themes during the congress. Eachday one of the keynote themes formed the basis for the day'sscientific sessions and presentations.

TheInternationalCongress of

PlantPath'ology

(ICPP)

Theme 1: Plant Pathology in the AsiaJPacific region• The Asia/Pacific region is an important agricultural sector ofthe world. Climatic factors such as highly disease conduciverainfall and temperatures contribute to the development ofseveral plant diseases that lead to significant losses.

Theme 2:Towards Integrated management of soilbornediseases.• Soilborne diseases pose a significant threat to agricultureand forestry and as such can result in significant losses toproduction and revenue.

Theme 3: Host/Pathogen interactions and molecular plantPathology• Molecular studies of host/pathogen interactions allowscientists to understand the biology of many plant pathogens.This contributes to the development of efficient control andbreeding strategies against many economically importantpathogens.

13

Theme 4: Towards Integrated management of airbornediseases.• Airborne diseases are some of the most destructive diseasesthat often lead to epidemics. This is because large amounts ofpathogen inoculum are built up and dispersed over largedistances in a short time period. Rusts are especially wellknown for their air dispersal and here, Eucalyptus rust, poses aserious threat to plantation forestry worldwide.

Theme 5: Knowledge transfer for Plant Pathology.• To control a plant disease effectively, sufficient andcompetent transfer of knowledge between plant pathologists iscrucial. Of even greater importance is the transfer of thisknowledge, and solutions, to farmers and foresters.

Various aspects of the research that is conducted in FABI andthe TPCP were presented at the conference. These includedposters on diseases such as Mycosphaerella, Cryphonectria,Diplodia, Ceratocystis, Fusarium, pink disease and bacterialblight. Professor Mike Wingfield delivered the DanielMcAlpine memorial lecture with a paper entitled "Exoticplantation forests in the Southern Hemisphere are increasinglythreatened by diseases". This seminar focussed onCryphonectria canker of Eucalyptus and the research that hasbeen conducted by the TPCP in South Africa. ProfessorBrenda Wingfield presented a paper entitled "Understandingthe threat of pitch canker to exotic plantation forestry in theSouthern Hemisphere". This paper focussed on the problem

of this disease in South African nurseries, difficulties that thepitch canker fungus causes at establishment and variousmethods of identification that have been used in diagnosingthe disease. These two seminars were among severalpresentations and posters that focussed on forest pathology.These papers generated much interest and lively debate amongthe delegates.

Apart from the many scientific sessions at the congress, therewas a full social itinerary for the delegates as well as a numberof field tours. On the first evening of the congress delegatesattended a welcoming cocktail evening with wine and verygood edible mushroom treats, allowing delegates to reunitewith old friends and make new bonds with other scientists.Congress delegates were also treated to a Barbeque evening atone of the local vineyards where they indulged in more goodfood and local wine. To end the congress, a Pacifica dinnerwas held where delegates could enjoy traditional foodaccompanied by dance and music from the New ZealandMauri culture. Following the congress, Mike Wing field andJolanda Roux participated in a two-day forest pathology fieldtour on the North Island. They looked at several importantdiseases including Dothistroma needle blight, Armillaria rootrot of Pines, Mycosphaerella Leaf Blight of Eucalyptus andSeridium canker of Cypress.

The ICPP congress was a great success. Much was learnt byall the attending delegates, and many new contacts were made.

&[]3@u~©u@ [pm@GYXJU[}{]~ m~©~~U

D~u~m~&uD@~&[s ©@~@m~@@ [p@m

[P[S&~U [P&U[}{]@[S@@W [}{]~[S[Q)D~

©[}{]mD@u©[}{]Mm©[}{]~ ~~W ~~&[S&~[Q)

Taxonomic re-evaluation of the generaCryphonectria and Endothia based onmorphology and DNA sequence data.

Henrietta Myburg, Marieka Gryzenhout, Brenda D. Wingfield,R. Jay Stipes and Michael 1. Wingfield

The fungal genera Cryphonectria and Endothia includesome of the most important pathogens of forest trees.Despite their importance, their generic and species leveltaxonomy has never been investigated at the DNA level.The objective of this study· was to comparerepresentative species of Cryphonectria and Endothia,for which cultures were available. Variation insequences of the ITS 1 and ITS2 regions of the ribosomalRNA operon and two regions of the D-tubulin genewere considered. In addition, morphologicalcharacteristics of these species were studied. Twodiscrete clades representing Endothia and Cryphonectriawere strongly supported in the emerging phylogenetictrees. The Cryphonectria clade included C. parasitica,C. macrospora, C. nitschkei, C. eucalypti and C.radicalis. The genus Endothia was characterised only

by E. gyrosa isolates. Isolates representing E.viridistroma and E. singularis, grouped separately fromeach other and outside the Endothia clade. Isolates ofthe Eucalyptus pathogen, Cryphonectria cubensis, and asupposed Cryphonectria species from New Zealandformed two groups separate from Cryphonectria andEndothia. The different clades representing Endothia,Cryphonectria, C. cubensis, the New Zealand isolates,E. singularis and E. viridistroma could be distinguishedbased on anamorph and stromatal morphology as well asascospore septation. Results of this study show clearlythat Cryphonectria and Endothia are polyphyletic andthat their taxonomy must be revised.

Bacterial blight of Eucalyptus species in Uganda.Grace Nakabonge, Teresa Coutinho, Jolanda Roux and

Michael Wingfield

Over exploitation of natural forests has. significantlyincreased the dependency on exotic plantations for thesupply of wood and wood products in the tropics andSouthern Hemisphere. In Uganda, Eucalyptus species

14

are amongst the most widely grown plantation trees,covering 40% of afforested areas. Diseases, however,reduce their growth rate, survival and product quality.Most Eucalyptus diseases are caused by fungi, butbacterial pathogens are increasing in importance. In1999, a disease was reported in the Entebbe area alongthe shores of Lake Victoria. Diseased trees showed leafspots, lesions along the main leaf veins, prematureabscission of the leaves, die-back and death. Isolationswere made from wilted shoots and from leaves withwater soaked lesions, consistently yielding bacterialisolates. The samples were characterized using Gramstain, Hugh-Leifson oxidation fermentation, oxidase andcatalase tests as well as 16S Rrna gene sequences. Thebacterium was identified as Pantoea ananatis, ,a knownleaf pathogen of Eucalyptus spp. In South Africa.Bacterial blight is consequently of great concern interms of establishment of future plantations, especiallyas it affects young trees. In South Africa, it has beenshown that there is great variation in susceptibility ofdifferent Eucalyptus species and clones. To enhanceproducti vity, the Ugandan forestry industry should thusestablish trials to select bacterial blight tolerant plantingstock .. This study represents the first report of bacterialblight, caused by P. ananatis in a country other thanSouth Africa.

Rapid identification of African strains ofRalstonia solanacearum.

J.P. Weich, S. Poussier , D. Trigalet-Demery, A. Trigalet,G. Nakabonge, D.K. Berger and T.A. Coutinho

Ralstonia solanacearum, the causative agent of bacterialwilt, is one of two bacterial pathogens known to infectEucalyptus in South Africa. Rapidly and accuratelyidentifying the causative agent in a plantation iscrucially important to managing the disease. In thisstudy, a rapid identification method, developed inFrance, was used to confirm the identity of Ugandan andCongolese isolates thought to be R. solanacearum. Themethod used is a PCR-RFLP technique that is based onthe hrp (hypersensitive response and pathogenicity) generegion. This region is needed by many plant pathogenicbacteria to cause symptoms on susceptible hosts and ahypersensitive reaction on resistant or non-hosts. Threeprimer pairs were used. The restriction enzymes AvaI,BssHII, HaeII, and PvuII were used selectively: to cutthe PCR products obtained which yielded two to four

FABI becomingFabylon?A stranger walking through the passages of the FABImight wonder if he/she has not perhaps rediscovered theBabylon of old? This impression would arise due to the

bands in a distinct pattern. The relatedness amongisolates of R. solanacearum from Congo and SouthAfrica were also determined using AFLP's. The AFLPresults showed that the Congo isolates and two of theSouth African isolates grouped together, based on thepresence/absence of AFLP fragments. The AFLPgroupings also corresponded with biovar groups. Thediagnostic technique will now enable researchers toidentify the pathogen rapidly and management strategiescan then be implemented within a week after anoutbreak occurs. .

Population structure of Cryphonectria cubensisin South Africa.

R.N. Heath, 1. Roux, N.A. van der Merwe, B.D. Wingfield andM.J. Wingfield

Cryphonectria cubensis is a canker pathogen ofEucalyptus in tropical and sub-tropical regions world-wide. In addition to its presence on Eucalyptus, thispathogen has been recorded on clove and Tibouchinaspp. In South Africa, C. cubensis results in symptomson Eucalyptus, which are different to those elsewhere inthe world. Recently, analyses of DNA sequence datahave shown that the fungus is different to C. cubensisoccurring in South America, South East Asia andCentral Africa. The origin of the South African fungusknown as C. cubensis has thus been in question. Arecent survey of indigenous Myrtaceae in South Africa,led to the discovery of C. cubensis on native Syzigiumspp. Detailed morphological and molecularinvestigations confirmed that the fungus is identical tothe one found on Eucalyptus in South Africa. Theprimary aim of this study was to gain insight into thepopulation structure of this fungus, using vegetativecompatibility and amplified micro satellite markers.Results showed that the population from Syzigium spp.has greater genetic diversity than populations fromEucalyptus in South Africa. Isolates from Syzigium alsohad a lower level of linkage disequilibrium than apopulation isolated from South African Eucalyptus.Therefore, the Syzigium population displays randommating, which is not true for the Eucalyptus population.These observations provide good evidence that thefungus known as C. cubensis in South Africa, originatedfrom native Myrtaceae in this country and subsequentlybecame established on exotic Eucalyptus andTibouchina spp.

large number of foreign Post Doctoral fellows, Mastersand Ph. D. students, that populate FABI. Of course, themany South Africans speaking a wide array of locallanguages contribute to this, not unpleasant, cacophony.

15

An informal survey was conducted recently in FABI todetermine exactly how many languages are currentlyspoken in the Institute. Of the 66 people who respondedto the questionnaire, all but one FABIAN, speaks two or

more languages. And it is known that the one personwho recorded that she only speaks English, understandsAfrikaans perfectly well! The person who speaks themost languages in FABI, is a South African Ph. D.student, Bongani Maseko, who speaks nine languages.Then there are seven FABIANS who each speak sixlanguages, five who speak two languages, four whospeak four languages, twelve speaking three, and theremaining 34 speaking two languages each.

The languages spoken (numbers of people in brackets),from South Africa included: Afrikaans (30), Zulu (5),Xhosa (4), S-Sotho (4), Tswana (4), N-Sotho (3), Swazi(2), Ndebele (1), and Hindi (1). The number ofAfrikaans speaking people seems high, but this is mainlybecause almost all the African language speakers alsospeak Afrikaans as a second or third language, while

Graduations

very few of the Afrikaans speaking people speaks any ofthe African languages. FABIANS from the remainderof Africa speak the following languages: Arnharic (3),Orornigna (2), Swahili (1), Luganda (1), Shona 0),Tonga (1), Chewa (1), Ndebele (1), Herero (1). Thosefrom Europe and Asia: German (7), French (3),Azerbaijanian (1), Russian (1), Turkish (1), Serbian (1),Portuguese (1), Italian (1), Norwegian (1) and Chinese(Mandarin) (1). Then there is 5 FABIANS from SouthAmerica who speaks Spanish. All of the respondentsspeak English, which is also the operational language inFABI.

In celebration of the international flavour in FABI, theFABI Christmas card for the past festive season,featured the season's greetings in 22 of the 29 languagesspoken in this very mnlticultural.institute.

We wish to congratulate the following people who recently received their degrees.

Grace Nakabonge - M.Sc. Cum Laude - Diseases associated with plantation forestry in Uganda.

Rodrigo Ahumada - M.Sc. Cum Laude - Diseases in commercial Eucalyptus plantations in Chile, withspecial reference to Mycosphaerella and Botryosphaeria species.

Mohammed Abdo - B.Sc. honours - Rhizobium spp. associated with various hosts in South Africa includingAcacia mearnsii.

Prem Govender - Ph.D - Soil invertebrate pests in the re-establishment of plantations in South Africa.

Xudong Zhou - Ph.D - Taxonomy and biology of ophiostomatoid fungi associated with coniferrinfestin barkbeetles.

IMPORTANT: PLEASE READ THIS

In order for us to coordinate our services to· you please help us by using the following contactaddress:

POSTAL ADDRESS STREET ADDRESS FOR DELIVERIES

Tree Pathology Co-operative ProgrammeAtt: Prof Michael J WingfieldMondi Professor of Forest Pathology and Directorof FABIForestry and Agricultural Biotechnology Institute(FABI) University of PretoriaPretoria 0002

FABILunnon RoadUniversity of PretoriaHillcrestPretoria

Tel: 012-420 3939 Fax: 012 420 3960 e-mail: mike.wingfield@fabi.up.ac.za http//www.up.ac.za/academic/fabi/tpcp

Editor: Jolanda Roux Design and Layout: Susan Christie16