fruit crop improvement
TRANSCRIPT
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Credit Seminar (Hort 591) Date: 26-10-16
Fruit crop improvement through tissue culture techniques- Achievements and Future
prospects
Kuldeep PandeyL-2015-A-85-M
Department of Fruit SciencePunjab Agricultural University, Ludhiana
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Plant Tissue Culture
Plant tissue culture refers to the cultivation of plant organs, tissue or cells in vessels using artificial media
Also referred to as cell, in vitro, axenic or sterile culture
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The theoretical basis for plant tissue culture was proposed by Gottlieb Haberlandt in his address to the German Academy of Science in 1902
Shoot bud regeneration from Tobacco suspension culture was reported by White in 1939
First plant from the mature plant cell was regenerated by Braun in 1959
Most of the modern tissue culture media have been derived from the work of Skoog and coworkers during 1950s and 1960s
History of Plant Tissue Culture
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The first embryo culture was done by Hanning in 1904 who cultured mature embryo of some crucifers
This technique was soon applied by Laibach in 1925 recover hybrid progeny from interspecific cross of in Linum
Haploid plants from pollen grains were first produced by Maheswari and Ghua in 1964 by culturing anthers of Datura
In 1960, Cocking produced large quantities of protoplast by using cell wall degrading enzyme
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Application of Tissue culture techniques in Fruit crop improvement
Recovery of virus free plant
Somatic embryogenesis
Somaclonal variation
Embryo culture
Anther culture
Somatic hybridization
Cryopreservation
Micro-grafting
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Recovery of virus free plant
Viruses are eliminated by:
A.Thermotherapy
B.Cryotherapy
C.Chemotherapy
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Species Virus Therapy ReferenceP. dulcis cv. Labanese
Candidatusphytoplasma
In vitro thermotherapy Chalak et al(2005)
P. armeniaca PPV, PDV, PNRSV
In vitro thermotherapy Laimer et al(2006)
P. ameniaca cv. Bebecou
PPV In vivo and in vitro thermotherapyshoot tip culture
Koubouris et al(2007)
P. avium PNRSV, PDV In vivo thermotherapy Howell et al(2001)
P. avium cv. Early River
ACLSV, PNRSV, PDV
In vitro thermo, chemotherapy
Cies´lin´ska (2007)
P. domesica cv. CacanskaLepotica
PPV In vivo chemotherapy Paunovic et al(2007)
P. domesica cv. Earliblue
PNRSV In vitro thermotherapy Dziedzic (2008)
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Plant species Virus eliminatedAnnanas sativus (Pineapple) Unspecified Fragaria sp (Strawberry) Complex of viruses, crinkle, edge, vein
banding, yellow virus complex, pallidosis
Malus sp (Apple) Latent viruses , Apple chlorotic leaf spot virus
Musa sp (Banana) Cauliflower mosaic virus, Bunchy top virus, Musa mosaic virus
Rubus ideaus (Raspberry) Mosaic
Vitis vinifera (Grape) Grapevine fan leaf virus, Arabis mosaic virus
Important Fruit Crops in which virus elimination has been successful using meristem culture
(Khurana et al 2003)
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Somatic Embryogenesis
Somatic embryogenesis has four well recognized phases:
(i)Induction(ii)Development(iii)Maturation(iv)Germination
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Somatic embryogenesis and plant regeneration from immature embryos of Carica papaya x Carica cauliflora cultured in vitro
Fig- Morphology and isozymes of Carica papaya, C. cauliflora, and interspecific hybrids of these two species derived from embryo culture: (A) Hybrid plant(B) C. papaya plant(C) C. cauliflora plant(D) Leaves of (left to right) C. papaya, hybrid, and C. cauliflora.
(Chen et al 1999)
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Somatic embryogenesis and plant regeneration inPsidium guajava L. cv. Banarasi local
.
(Rai et al 2007)
(A) Zygotic embryo explant from 10-week-old fruit (B) Globular to cotyledonary stage somatic embryos (C) Maturation of somatic embryos (D) Initiation of germination of somatic embryos (E) Plantlets obtained by germination of somatic embryos (F) A healthy and well-hardened plant of guava
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Species Explant ReferenceP. mume cv. Nanko Immature
cotyledonTsukamoto et al(2007)
P. avium Zygotic andSomatic embryo
Garin et al(1997)
P. avium Zygotic embryo Talleux et al(1999)
P. cerasus Immaturecotyledon
Tang et al (2000)
P. avium Protoplast David et al (2006)
P. persica cv. Bell of Georgia, Bailey, Tennessee Natural, Nemaguard, Encore
Zygoticembryo
Srinivasan andScorza (2007)
P. incisa cv. February Pink Leaf, root Cheong andPooler (2004)
P. subhirtella autumn rosa Petiole Machado et al(1995)
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Cultivar(s) Explant Response PGRs Media Reference
Cambodiana, Carabao, M20222,Turpentine N2-1-4-3, Earlibush,Chino, Turpentine N2-1-7-2,Sabre and Ono
Ovules Somatic embryogeneis(SE) and precociousgermination
With or Without1–2 mg/l ; 20%coconut water
Modified MS medium+400 mg l−1 glutamine+100 mg l−1 ascorbic acid
Litz et al(1982)
Amrapali Nucellus SE and germination
5–20 μM BAP MS and B5 media
Laxmi et al(1999)
Carabao and Manila Super
Nucellus SE Modified Medium
Patena et al(2002)
Ambalavi Nucellus SE and plantletregeneration
0.25 mg/l BAP+ 1.0 mg/l NAA;0.5 mg/l 2ip
Modified medium (BM)
Chaturvediet al (2004)
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Important fruit crops in which protocols are being standardized for efficient cloning of plants through
somatic embryogensisMango (Mangifera indica)Banana (Musa sp.)Citrus (Citrus sp.) Grapes (Vitis venifera) Pineapple(Annanas sp.)Papaya (Carica papaya)PomegranateLitchi (Litchi chinensis)Apple(Malus domestica)Peach(Pyrus sp)Strawberry(Fragaria sp.)Datepalm (Phonix dectylifera)Walnut (Jaglans regia)Ber (Ziziphus sp.) (Singh 2011)
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Somaclonal variation
Somaclonal variants may arise due to:
1.Chromosomal structural changes2.Gene mutation3.Plasma gene mutation4.Gene amplification5.Karyotype changes6.Mitotic crossing over etc.
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Application of Somaclonal variation1.Novel variants – Thornless Blackberry released in New Zealand as cv Lincoln Logan Hall et al (1986)
2.Disease resistance- Banana- F. oxysporium sp.cubens Apple- Phytopthora cactorum Peach- Xanthomonas comp.pv.pruni Pseudomonas syringae pv syringae3.Abiotic stress resistance a)Cold tolerance b)Salt tolerance c)Drought tolerance d) Aluminum tolerance
4.Herbicide resistance5.Insect resistance6.Seed quality
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Cavendish banana cultivars resistant to Fusarium wilt acquired through Somaclonal variation in Taiwan
(Hwang et al 2004)
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Name of the somaclonal variant
Cultivar from which derived
Usefulness Reference
CIEN-BTA-03 Cavendish Resistant to yellow Sigatoka
Trujillo and Garcia 1996
TC1-229 Tai-Chiao No. 1 Semi-dwarf and resistant to Fusarium wilt
Tang et al 2000
Tai-Chiao No. 1
Formosana
Giant Cavendish
Giant Cavendish
Resistant to Fusarium wilt
Resistant to Fusarium oxysporum f. sp. cubense (Foc)race 4; bunch 40% heavier
Hwang 2002
Hwang 2002
TC2-425 Tai-Chiao No. 2 Larger bunch size Hwang 2002
GC-138 Pei-Chiao Growing cycle 1–1.5 month shorter
Hwang 2002
------ Grand Naine Dwarf with superior quality
Stover 1987
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Somaclonal Variation: a Useful Tool to Improve Disease Resistance of Pear Rootstock ‘Old Home × Farmingdale’ (OHF
333) (Pyrus communis L.)
Nacheva et al (2014)
Class 0 – plants without visual symptoms of the infection;
Class 1 – plants in which the leaf cut has blackened;
Class 2 – plants in which the cut and the central nerve have blackened
Class 3 – plants in which the leaf and the leaf petiole have blackened
Class 4 – plants in which the apical shoot and the upper part of the plant have blackened
Class 5 – the whole plant is dying
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Somaclone Degree of susceptibility
Somaclone Degree of susceptibility
Control 5 X-16 0
VIII-1 0 X-8 1
X-1 0 VI-2-1 1
VI-2 0 VI-3-1 0
VI-3 4 X-1-3 0
X-1-1 0 VIII-1-1 4
VI-1 0 X-1-4 0
VI-8 1 X-1-5 2
VIII-5 0 VI-1-1 4
VI-12 4 VI-3-2 2
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Studies on Somaclonal Variation in Strawberry (Fragaria x ananassa Duch.) Cultivars
(Kaushal et al 2004)
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Embryo CultureApplications of embryo culture:
1.Recovery of Distant Hybrids
2. Recovery of Haploids plants from interspecific crosses
3. Propagation of orchid
4. Shortening the breeding cycle
5. Overcoming dormancy
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Some fruit Plants in where embryo culture used- Apple, Cherry, Pear, Peach, Grapes, Banana, Papaya (Singh 2005)
The first peach embryo culture studies (Davidson 1933, 1934)
i.Shortening the breeding cycle of peachii.Increasing germination of embryos from early ripening cultivarsiii.From interspecific crosses
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Stage of fruit Media Reference
Immature fruitlets (35–45 days old)
MS basal medium(9 mg/l BA + 3 mg/l kinetin + 400 mg/l glutamine + 500 mg/l activated charcoal + 60 g/l sucrose
(Nathhang 1999). Chandra et al (2003)
Fruitlets at 6–8 weeks after pollination
MS medium(casein hydrolysate(1.25 g/l) and sucrose (4.5%).
Sahijram et al (2005)
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Fig- Fruits of:(A)Seeded berries from C41-7(B)Parthenocarpic berries from C41-7(C)Mature berries from C41-5(D)Overripe berries from C41-5.
A sternosprmocapic seed less Vitis vinifera x Vitis routundifolia Hybrid Developed by
Embryo Rescue
Ramming et al 2000
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Fig- Effect of maturity status and medium composition on callus induction in zygotic embryos of Pisang Jajee
Embryo rescue and plant regeneration in banana (Musa spp.)
(Uma et al 2010)
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Fig-Effect of embryo maturity and media composition on plantlet regeneration from callus tissue derived from zygotic embryos
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Anther/Pollen Culture
Application of anther culture:
1.Obtaining haploids2.Obtaining homozygous diploids
Regeneration of androgenic embryos in apple (Malus x domestica Borkh.) via anther and microspore culture.
(Hofer 2005)
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CitrusMalus domestica Pyrus communisPyrus pyrifoliaPrunus persica Prunus aviumPrunus domestica Prunus armeniaca Vitis vinifera
Actinidia deliciosaOlea europaeaMorus alba Musa balbisiana (BB)Carica papaya Annona squamosa FeijoasellowianaOpuntia ficus-indicaEriobotrya japonica
Doubled haploid production in fruit crops
(Germana 2006)
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Androgenic potential in coconut (Cocos nucifera L.)
(Perera et al 2008)
Fig- (a)Isolated rachilla (r) containing male (mf) and female flowers (ff)
(b) Close view rachilla
(c) Isolated male flower: Anthers are totally covered by the petals(p)
(d)Anthers (an) attached to the filaments (f)
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Fig-(a) Embryos (em) derived from anther (an); (b) Mature embryo with germination point (gp); (c) Germinating embryo with a shoot (s) emerging by splitting of haustorium (h); (d) Calli (ca) bearing anther; (e) An anther bearing both callus (ca) and embryos (em); (f) Somatic embryos (se) developed from anther derived callus; (g) A complete plantlet derived from anther
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Somatic Hybridization
The technique of somatic hybridization involves the following four steps:(i)Isolation of protoplast(ii)Fusion of protoplast(iii)Selection of hybrid cells(iv)Regeneration of hybrid plants
Applications of somatic hybridization
1.Symmetric hybrid2.Cybrid
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(Cybrids) regenerated via symmetric protoplast fusion in Citrus
Fusion combination References
C. sinensis cv. Bonanza navel + C. sinensis cv. Valencia
Deng et al (2000)
C. sinensis cv. Newhall navel + C. sinensis cv. Anliu Deng et al (2000)
C. reticulata cv. Red tangerine + Yuzu (C. junos) Deng et al (2000)
Page tangelo + C. sinensis cv. Valencia Deng et al (2000)
Page tangelo + C. sinensis cv. Gailiang Deng et al (2000)
Swinglea glutinosaa + Sour orange (C. aurantium) Moreira et al (2000)
C. sinensis cv. Rohde Red Valencia’ + C. reticulata cv. Dancy
Moreira et al (2000)
C. reticulata cv. Cleopatra + Sour orange (C. aurantium) Moreira et al (2000)
Microcitrus papuana + Sour orange (C. aurantium) Xu et al (2004)
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Fusion combination References
C. reticulata.cv. Shekwasha + C.sinensis cv. Valencia Guo and Grosser (2005)
C. sinensis cv. Salustiana + C. aurantifolia cv. Mexican lime
Fuster et al (2005)
Page tangelo + Rough lemon (C. jambhiri) Guo et al (2006) C. reticulata cv. Chios mandarin + C-35 citrange (C.sinensis× P. trifoliata)
Guo et al (2006)
C. reticulata cv. Chios mandarin + C-35 citrange (C.sinensis× P. trifoliata)
Dambier et al (2011)
C. reticulata cv. Chios + 4475 citrumelo (C.paradisi× P. trifoliata)
Dambier et al (2011)
C. reticulata cv. Chios + citrandarin (C. reticulata × P. trifoliata)
Dambier et al (2011)
C.aurantifolia cv. Macrophylla + Carrizo citrange (C.sinensis × P.trifoliata)
Dambier et al (2011)
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Fusion combination References
Tommy Atkins+ Kensington Pride Rezazadeh et al (2011)
Keitt + Kensington Pride Rezazadeh et al (2011)
Haden + Kensington Pride Rezazadeh et al (2011)
wild pear + Colt cherry Ochatt et al (1989)
Rubus ideaus + R. fruticosus Mezzetti et al (2001)
Diospyrus glandulosa +D. kaki(persimmon)
Tamura et al (2002)
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Synthetic Seed
The concept of synthetic seed was given by Murashige (1977), but first report on the development of synthetic seeds was published by Kitto and Janick (1982).
There are two types of synthetic seeds: HydratedDesiccated
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Scope of Synthetic Seeds
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Plant species Propagules encapsulated References
Actinidia deliciosa (Kiwifruit)
Apical and axillary buds Adriani et al (2000), Romay Alvarez et al (2002)
Ananas comosus (Pine apple)
Axillary buds Soneji et al (2002), Gangopadhyay et al (2005)
Carica papaya (Papaya) Somatic embryos Castillo et al (1998)
Citrus sp. Somatic embryos Antonietta et al (1999, 2007)
Malus pumila (Apple rootstock M.26)
Apical and axillary buds, Nodes, Shoot tips
Piccioni and Standardi (1995), Piccioni (1997), Capuano et al (1998),Sicurani et al (2001), Brischia et al (2002)
Mangifera indica (Mango) Somatic embryos Ara et al (1999)
Musa sp. (Banana) Shoot tips, Shoot apices, Somatic embryos
Ganapathi et al (1992), Rao et al (1993), Suprasanna et al (2001),Ganapathi et al (2001)
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Plant species Propagules encapsulated
References
Psidium guajava (Guava)
Somatic embryos, Shoot tips, Nodal segments
Akhtar (1997), Rai and Jaiswal (2008), Rai et al (2008)
Punica granatum (Pomegranate)
Nodal segments Naik and Chand (2006)
Pyrus communis (Pear) Shoot tips Nower et al (2007)
Pistacia vera (Pistachio)
Somatic embryos and embryogenic mass
Onay et al. (1996)
Vitis vinifera (Grape) Somatic embryos Das et al (2006)
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CryopreservationCryopreservation can also be employed for uses other than germplasm conservation, such as cryoselection i.e., the selection through freezing of samples with special properties or cryotherapy i.e., the elimination of viruses from infected plants through apex cryopreservation.
Effect of cryopreservation on the sanitary state of a cv. Prunus rootstockexperimentally contaminated with Plum pox virus.
(Brison et al 1997)
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Species Explant Reference
P. dulcis cv. Ferragnes, Supernova
Cell suspension Damiano et al (2003)
P. armeniaca Dormant bud Irina et al (2006)
P. avium Embryonic tissue March(2005)
P. avium In vitro shoot tip Shatnawi et al (2007)
P. avium Seed Chmielarz (2009)
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Elimination of grapevine virus A (GVA) by cryopreservation of in
vitro-grown shoot tips of Vitis vinifera L
(Wang et al 2003)
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Low temperature storage of mango (Mangifera indica L.) pollen
(Dutta et al 2013)
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Micro-grafting
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(Naz et al 2007)
In vitro studies on micrografting technique in two cultivars of Citrus to produce virus free plants
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Micrografting and ex vitro grafting for somatic embryo rescue and plant recovery in avocado
(Persea americana)
(Raharjo et al 2005)
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Micrografting of almond (Prunus dulcis Mill.) cultivars “Ferragnes” and “Ferraduel”
(Yıldırım et al 2010)
Method of micrografting
Success of micrografts (%)“Ferragnes” “Ferraduel”
Vertical slit 90.0 100.0
Wedge on the stump
60.0 70.0
Effects of micro grafting method on success of micrograft
Effects of scion size on the success of almond micrografting
Size of microscion (mm)
Success of micrografts (%)“Ferragnes” “Ferraduel”
4 93.0 100.0
815
100.093.0
100.0100..
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Species Virus Therapy ReferenceP. dulcis cv. Marcona, Fournat,uono, Ferraduel
PNRSV, PDV In vitro micrografting
Rizqi et al(2001)
P. persica PRSV, PDV, CLSV
In vitro shoot tip grafting
Navarro et al(1982)
P. persicacv. Suncrest
PNRSV In vitro micrografting
Heuss-LaRosaet al (1995)
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Future Prospect of Tissue culture in Fruit crop breeding
Future prospects of tissue culture in fruit crop breeding appears to galore
i. In present scenario, where the growers face the problem of shortage of disease free planting material, the tissue culture is a potential means to tackle this problem
ii. Inoculation of AMF or other beneficial microbes during weaning
iii. Expanding the existing genetic resource in fruit crops
iv. Conservation of wild species
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