Production and Characterization of Haploids in Brinjal (Solanum melongena L.)
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Date
2022
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Punjab Agricultural University
Abstract
A study was conducted for production and characterization of haploids in brinjal. Induction of haploids in brinjal was attempted through two methods androgenesis and interspecific hybridization. For microspore androgenesis among donor hybrids, PSB-7-2 X P-219 had the highest (14.6 in 63 days) number of calli per ml of microspore suspension, followed by SR-5 x SR-9322 (4.4in 69 days) and SC-15-2 x MR-319 (2.1 in 73 days). Overall, NLN medium supplemented with 2 mg/l 2, 4-D + 20g/l Sucrose induced the maximum number of calli per ml of suspension (13.3 calli in 62 days). In this study, four days temperature pretreatment (35̊ C) in distilled water induced the uppermost number of calli per ml of suspension (11.2 calli in 66 days). The embryogenesis (%) of microspore derived calli was the most effective in MS + 4 mg/l Zeatin + 0.2 mg/l IAA+ 20 g/l of sucrose + 8g/l agar (R1) as compared to MS + 2 mg/l Zeatin + 0.2 mg/l IAA+ 20 g/l of sucrose + 8g/l agar (R2) for all the donor hybrids. In total, 260 plants were rooted on half-strength MS medium and only 65 (25.0 %) could be established as complete plants after hardening. In the confirmation, the molecular analysis with three SSR markers established complete homozygosity (microspore origin) in all the plants developed from two donors. Chromosome counting (n=12) at diakinensis or metaphase stage of meiosis in pollen mother cells categorized 65 regenerants into 5 haploids, 30 doubled-haploids, 29 mixoploids, and one triploid. Many chromosomal abnormalities in PMCs also affcetd the reproductive features of heteroploids. In anter androgenesis of donor hybrids, SC-15-2 x MR-319 and SR-5 x SR-9322 were at par with each other for percentage of calli formation from expanded anthers with values 73.4 % and 71.3 % respectively. Donor hybrid SR-5 x SR-9322 also showed direct embryo regeneration from expanded anthers with value 2.8 %. Among different media combinations, the maximum calli formation from expanded anthers with in minimum days was observed in MS + 2 mg/l IAA + 0.5 mg/l Kinetin (65.8 % and 28.5 days). Direct embryo regeneration from expanded anthers was observed only in media composition MS + 2 mg/l IAA + 0.5 mg/l Kinetin (1.6 %). However, the highest calli regeneration was observed in donor hybrid SR-5 x SR-9322 (15.5%) with in minimum days (21.7 days). Among media combinations, MS + 0.2 mg/l IAA + 4 mg/l Zeatin (19.5 %) had the maximum calli regeneration in the minimum number of days (21.4 days). Plants obtained through anther androgenesis via direct embryos regeneration from expanded anthers in SR-5 x SR-9322 at media combination MS + 2 mg/l IAA + 0.5 mg/l Kinetin also confirmed their genetic origin. Morphological observations and molecular marker EEMS 37 confirmed the possibility of the haploid nature of these plants. In wide hybridization, no chromosome elimination was observed in plants. In conclusion, microspore culture of F1 donor was the best approach to develop homozygous lines of genetic origin and considerable morphological and genetic variability.
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Jasmeen Kaur (2022). Production and Characterization of Haploids in Brinjal (Solanum melongena L.) (Unpublished Ph.D. Dissertation). Punjab Agricultural University, Ludhiana, Punjab, India.