Breeding for seedlessness in horticulture crops.pptx

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1. ITM UNIVERSITY GWALIOR PRESENTED BY: HIYA DASHORA MHVN1AG23003 M.Sc Horticulture {Vegetable S cience } BREEDING FOR SEEDLESSNESS IN HORTICULTURE CROPS SCHOOL OF AGRICULTURE GUIDED BY: Prof. (Dr.) C.K. Sharma Dr. Shubham Singh Rathore Department of Fruit science Master’s Seminar

2. Content Introduction Seedlessness trait and factors affecting it Characteristics of seedless fruit Parthenocarpy  Hormonal Regulation of Fruit Set Breeding methods Genome editing technology--- CRISPR- Cas Genetic Engineering Strategies for Seedlessness Breeding Recent Research Conclusion References .

3. Introduction A plant is considered to be seedless when its fruits are completely devoid of seeds, contain a much-reduced number of seeds or present aborted seeds. Seedless are obtained through parthenocarpy if the fruits develop without fertilization and by stenospermocarpy if the seeds abort after fertilization  Factors affecting Seedlessness or Factors inducing Seedlessness include certain environmental conditions, such as low or high temperatures, chemical treatments, chromosomal aberrations, and genetic factors.  Seedless fruit can be obtained through parthenocarpy if the fruits develop without fertilization and by stenospermocarpy if the seeds abort after fertilization

4. Seedless fruits are generally smaller in size compared to their seeded ones. B. In seedless fruits, other forms like reduced seed number, soft seed, hollow seed (Ex. grapes). C. Seedless fruits may result in thickening of pedicels in some cases (EX. Pear) D. Fruit surface is normally very smooth in case of seedless fruits compared to seeded ones except in case of some where the surface is warty . (EX. Guava) E. Seedless fruits in general mature later than their seeded ones. Characteristics of Seedless Fruit T he Pharma Innovation Journal

5. Parthenocarpy Greek word , literally means “ Parthenos : virgin karpos : fruit” is the natural, artificially induced, or genetically modified production of fruit without fertilization. In the absence of pollination, parthenocarpic plants will set seedless fruit. ( Gustafson, 1942 ) and According to Winkler (1907), Parthenocarpy : Primary Requirement to Produce Seedless Fruit . In artificial it is made between vegetative parthenocarpy, which allows fruit set without pollination, and stimulatative parthenocarpy, in which fruit set follows pollination, but subsequent fertilization is prevented, resulting in the production of seedless fruit . In genetic/natural distinction is often made between obligatory parthenocarpy (Ivy gourd), which always results in seedless fruit, and facultative parthenocarpy, which results in seedless fruit only when pollination is prevented . ( Eg : Tomato, brinjal and cucumber) Seedlessness is appreciated by consumers both in fruits for fresh consumption (e.g., grape, citrus, and banana) as well as in conserved or processed fruits (e.g., frozen eggplant, tomato sauce

6.  Seedlessness can contribute to increase the quality of the fruits when seeds are hard or have a bad taste. In the case of eggplant, the absence of seeds prevents browning and texture reduction of the pulp. Furthermore, seeds can produce substances that accelerate the deterioration of the fruit, as in watermelon and eggplant. In this regard, the absence of seeds can increase the shelf life of the fruits, allowing a better conservation. Continue http://www.jstor.org/stable/4353278 The mean temperature during the pre- flowering, flowering and fruit-setting period is the primary factor responsible for the production of seedless fruits.

7. Seedless okra production by indole 3-acetic acid micro syringe injection on flower bud, ovary and shoot xylem and its vitamin and mineral content development Scientia Horticulturae Pusa Bedana:It is seedless watermelon developed at IARI, by crossing Tetra 2 ( tetraploid ) x Pusa Rasal Cucumis sativus (Cucumber). Noll observed that the varieties Gurke and Grosse produced seedless fruits when pollination was prevented. In some the ovules made a little growth but no embryos were observed Luffa cylindrica. Scientist found that when treated the pistils of flowers that had just opened with a 1.0%o indole acetic acid-lanolin paste he was able to produce seedless fruits that were nearly as large as the seeded fruits

8.   Hormonal Regulation of Fruit Set A widespread agriculture practice for the production of seedless parthenocarpic fruit consists in treating flowers with phytohormones before pollination. Auxin, Gibberellins Cytokines Mixtures of these hormones have all been proven to be effective in inducing fruit development in the absence of fertilization in several crop species, for example vegetables like tomato and eggplants. Brinjal: Muthukrishnan and Srinivasan, 1980 Cucumber : Choudury and Phatak, 1960; Kim et al ., 1994 In crops such brinjal and cucumber, there is evidence that parthenocarpic fruit set is due high levels of endogenous auxin and GAs in the ovary. To improve fruit set citrus cultivars Auxin levels peak sharply 10 d after anthesis, and exogenous GA is essential for fruit development and the prevention of fruit abscission.

9. Fig : (a) different steps in fruit development, (b) t steps in seed development and (c) examples of seedless fruit. Varoquaux et al ., 2000

10. Breeding efforts for high-quality seedless citrus fruit have increased during the past few decades and few cultivars developed are : Satsuma mandarin ( Citrus unshiu  Marc.) Tahiti lime ( Citrus latifolia) Washington Navel orange ( C. sinensis ) Seedlessness in Parthenocarpic Citrus Cultivars Fig: Satsuma mandarin Fig: Tahiti lime Another approach for seedlessness involves the coupling of parthenocarpy with male sterility . Ploidy and Cytoplasmic Male Sterility Washington Navel orange

11. Ploidy breeding

12. 1. Triploid in Watermelon: Seedless watermelons are gaining popularity as the mature seedless melons become more available in the general market. 2. Triploid breeding in grape Production of large seedless berries with high quality is one of the most important objectives in table grape breeding programs Ploidy manipulation using crosses between diploids and tetraploids have yielded several valuable triploid cultivars in many of crops. Kihara , 1951 a.Marvel seedless b.Muscat alexandria c.Black king from champbell

13. 5. Mutation Breeding: Irradiation is a valuable tool to obtain seedless cultivars from seeded ones. Mutation affects pollen viability. Female and male sterility seem to be directly related, In Citrus spp with a lower seed number present a lower pollen viability, and chromosomes aberration during meiosis caused by irradiation are responsible for sterility. 3. climacteric seedlessness is good among fruits and have good traits like high protein content, high moisture content 4. Endosperm culture It is widely known as the fusion of three haploid nuclei. This is a good potential area to isolate triploid plants directly from the embryonic stage and cut short the lengthy process of growing and crossing which could develop into a possible seedless plant. Influence of Gamma Irradiation on Seedless

14. FIG: Visual scheme of gaining the seedlessness trait using (CRISPR/ Cas ) strategy Enormous way and endless possiblities Success rate is very high Open up new doors

15. In Many citrus cultivars the first approach is male sterility. could potentially set seedless fruit in the absence of pollination However, this approach is restricted to certain conditions under which male sterile citrus plants are isolated from male fertile plants. The second approach is seed coat destruction to obtain stenospermocarpy through the expression of targeted suicide genes during early seed coat development. In this case, parthenocarpy is not required for seedlessness. The third approach is female sterility. Destruction of the ovule or the stigma by suicide genes could trigger seedless fruit set as fertilization would be impossible. However, this approach is limited to cultivars with strong parthenocarpic traits . The fourth and last approach is increased expression or increased sensitivity to hormonal in the ovary or ovule. This approach, via genetic engineering of auxin expression or sensitivity, has been used successfully in several field crops Source: DOI:  https://doi.org/10.21273/JASHS.133.1.117

16. [A] Based on large numbers of experimental studies, Li et al. (2001) concluded that the main factors determining the success of embryo rescue are the sampling time, the parental genotype, the culture medium, and conditions. [B] The emergence of molecular markers further sped up the breeding process for seedless grapes. [C] Seedlessness evolution caused majorly due to spontaneous mutation in nature. D] Genetic makeup: Seedless fruits is a highly desirable trait and is due to natural causes and demand . [E] Induced mutations and Isolation of natural bud [F] Climatic conditions (Fig) , Location (Latitudinal changes- E.g.: Grapes ).  Seedlessness remains a main focus of breeders

17. Lack of availability morphological marker for adjudging seedlings at the early age In conventional breeding, only seeded genotypes can be used as seed parents, hence there is lot of limitations. When seedless genotypes are used as male parents, the pollen quality, viability etc. vary considerably and hence the results are not consistent . The average frequency of seedless progeny that has involve seedless parent can only give rise to 10-15% (on avg.) seedless progeny which is further reduced by poor germination. Introgression breeding to transfer quality and seedlessness and an alternative approach involving different crosses is quite challengeing In many population the time for evolving seedless hybrid is reduced to 5-6 years compared to 12- 15 years under the conventional system. Challenges faced by breeder

18. Advantages of Seedlessness Year-round production Gustatory advantage processing industry Seed cavity can be filled with fruit tissue (seedless pickled gherkin, which is more crunchy, firmer and fleshier than its seeded variety) Seedless watermelons develop a meaty texture and become overripe significantly later than seeded varieties. Lack of stability and uniformity in the expression of Parthenocarpy. Malformed fruits, Small sized fruits Hampers the production of commercial seeds Seedlessness results in hampered seed production and there is no room for the commercial seed production Limitation of seedless fruits

19. Recent research (December 2021) It is shown that, parthenocarpy occurs more in plurispermic (multi seeded) species than monospermic (single-seeded) (Picarella and Mazzucato, 2019) It is also reported that this phenomenon occurs more in cultivated species than in wild species

20. 1. Dokoozlian , N. 1994. Crimson Seedless research update: Reducing berry set and increasing berry size with gibberellic acid. Vine Lines ( Nwsltr . Univ. Calif. Coop. Ext. Fresno County) April. p. 1–3. 2.Li, G.; Wang, Y.; Tang, D.; Wang, X.; Luo , Q. The studies on embryo rescue techniques of ‘Thompson Seedless’ grape.  Plant Cell Tissue Organ  2001,  21 , 432–436 Li, J.; Wang, X.; Wang, X.; Wang, Y. Embryo rescue technique and its applications for seedless breeding in grape.  Plant Cell Tissue Organ Cult.  2015,  120 , 861–880.   3. Abbate L, Panno S, Mercati F, Davino S, Fatta Del Bosco S. Citrus rootstock breeding: response of four allotetraploid somatic hybrids to Citrus tristeza virus induced infection. European Journal of Plant Pathology. 2019;153(3):837-847. 4.Ahmed D, Curk F, Evrard JC, Froelicher Y, Ollitrault P. Preferential disomic segregation and C. micrantha /C. medica interspecific recombination in tetraploid ‘Giant Key’lime ; outlook for triploid lime breeding. Frontiers in plant science. 2020;11:939. 5Aleza P, Juárez J, Cuenca J, Ollitrault P, Navarro L. Extensive citrus triploid hybrid production by 2x × 4x sexual hybridizations and parent-effect on the length of the juvenile phase. Plant Cell Reports. 2012;31:1723- 1735. 6.Handaji Najat HB, Najat Arsalane HB. Selection of new promising seedless mandarins triploid hybrids from crosses between monoembryonic diploid female and diploid male parents. Research in Biotechnology. Food and Scientific Reports. 2017;1(4):25-27. 7. Hoshino Y, Miyashita T, Thomas TD: In vitro culture of endosperm and its application in plant breeding: approaches to polyploidy breeding. Scientia Horticulture. 2011;130:1-8. References