Androgenesis response studies in Guava (Psidium guajava L.)
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Date
2017
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Punjab Agricultural University, Ludhiana
Abstract
Development of haploids and doubled haploids through in vitro androgenesis greatly
accelerates the process of achieving homozygosity in crop plants including fruit trees. In tree
species including Citrus, Papaya, Neem, Mulberry, Custard apple and Apple, androgenesis have
been attempted successfully to overcome self in-compatibility barriers so as to raise
homozygous trees. In current study we attempted haploid production in three genotypes of
guava viz. Allahabad safeda, Purple Guava and Punjab Pink. Anthers with microspores at mid
to late uninucleate stage on N6 basal media containing 1% PVP showed maximum callus
induction of 16.3±0.27% in Allahabad Safeda followed by 10±0.41% in Punjab Pink and
4.6±0.27% in Purple Guava. The stereomicroscopic evaluation of callus indicated the callus
emergence from microspores. The callus induction frequency was enhanced from 1.7±0.27%
to 27.7±1.18% by cold (4 ºC) and mannitol (8%) pre-treatment for 3-7 days to unopened stage
IV flower buds. 77% calli survived on best regeneration media, nourished with liquid form in
the mother test tube without mechanical disturbance for 1 month. One callus exhibited
secondary structure on regeneration media. Isolated microspores showed increase in size on
MS and N6 media containing 2,4-D (2.5 mg/L) and Kinetin (2.5 mg/L) and few anomalies on
MS media fortified with BAP (0.2 mg/L) and Kinetin (0.2mg/L). 2/50 ovaries underwent
callusing after 4 months of in vitro culturing on 1% PVP containing basal MS media and callus
finally underwent necrosis after one week of sub culturing on MS media fortified with BAP
(0.2 mg/L) and Kinetin (0.2 mg/L). Haploid nature of callus can further be evaluated by
counting the chromosome number in the cells.
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