Improvement of bitter gourd (momordica charantia l.) Through breeding and biotechnology

Abstract

A field experiment was carried out in the Fruit Research Station, Bindpur, Rajshahi during 2001-2003 with 6-pasestal and 5-parental diallel sets to determine the gone effects for different quantitative traits te, vegetative-reproductive and al traits in bitter gourd (Momordica charanis L.) Significant differences were observed for most of the traits. The range of variation was enough for all the traits studied, which showed greater scope for their selection as breeding materials. Varians analysis of diallel exhibited the presence of additive gene effects for most of the traits, except BBB, FBN, FEN, and ETS, PTS, GTT-LP, respectively for vegetative-reproductive and anatomical traits of 5-puntal diallel. Out of eighty-nine traits (in both dillelsets), the item was significant for 83 traits, opt ID, FPL, OD and FV of 6-pereal diallel and ID and FBN of 5-parental diallel. In this investigation, relatively lager value of 'a' component in comparison to those of '&' for all the traits were observed. So, the additive component of variation was more important for all the traits in their inheritance. The additive component 'D' was highly significant for all the traits, except LB12 and DRMF of 6-gatal diallel. Whereas, in 5-perectal dialled for vegetative- reproductive traits out of 35 traits the significant 'D' value was observed only for five traits (e, I, OL, FL, FD and FP. In anatomical traits the significant 'D' was observed in seven traits only. The item H1 was significant for most of the traits. The item 'D' was slightly more predominant in most of the traits than dominant component H. The components H, and F were significant for most of the traits and the values were also higher than their "D" values, which showed the importance of non-additive inheritance. It was observed that the H1 values were smaller than H, in most of the traits. So, equal frequency was present in the parents. The positive sign of Fin most of the traits indicated the presence of dominant genes, but in most of the traits it was non-significant, which indicated that dominant properties of genes was not valid. For most of the traits, was non-significant, epistis was not present in the pants. In 6-parental diallel partial dominance was present in 19 out of 30 traits. Whereas, most of the traits (24) showed over dominance in 5-parental diallel. Most of the traits indicated asymmetry of distribution with+,- alleles. In this experiment most of the traits exhibited excess dominant alleles Le, dominant alleles were more frequent than recessive alleles in punts in both of the diallel se The presence of variability and non additive nature was noted for meat of the tits. The heritability in narrow sense ranged from 15.52 (LBL2) to 99.25% (FV) in 6- Parental diallel, 36.41 (DRB) to 69.35 (FV) and 1.40 (PTS) to 50.15% (OPT-IVB) respectively in vegetative-reproductive traits and anatomical traits in 5-allel The regression of Wr on Fr were unity in most of the traits and therefore, the additive dominance model was satisfactory. Non-allelic interaction was not observed in the parents Combining ability was observed in 39 traits in both the diallel sets, GCA effects of parents and SCA effects in direct and reciprocal crosses were significant with a higher magnitude of C of GCA variance indicating that inheritance of these traits were predominantly governed by additive and additive x additive epistatic components. Among 6-paremal diallel of model-l method-ll, the genotype BGGBI was found to be the best general combiner for the important traits I, DRFF, FL, FV, FW and FFW. The genotypes BGGB4, BGGB13 and BGGB14 were the best the general combiner for DRMF, FNFW and HF/W respectively. Similarly 6-perectal diallel of model-l method-I showed th BGGB12 was the best general combiner for DRME, DRFF, FV, FW FFW and FNFW. For 5-parental diallel, the best performing GCA effect was in BOGB17 for IL, DRB, DRMP, DRFF, FL, FFT. The important traits FW, FFW, SF and FP had no significant general combiner. The parent BGGB20 was the best combiner for most of the anatomical traits. In the 6-parental dialled of model-l method-1, the best crosses with high SCA effects were BOOB14 x BOGB4 for DRMF BGGB6 x BOGB1 for DRFF, BOGB14 BGGB13 for FL, BGGBS BGGB4 for FD, BGGB14 BGGBI3 for FV, BGGB14x BGGB13 for FW, BOGB14 BOGBI3 for FFW, BOGB12 BOGB6 for HF/W, Similarly for 6-pantal dallel of model-t method-II the SCA effects showed that the best specific combinations were BGGB13 BGGB6 for DRMF, BGGB6 BGGB4 for DRFF, BGGB14 BOOB12 for FL, BOOB13 B0084 for FD, BOGB13 BOGBI for FV, BOGB12 BOGBI for FW, BGGB12 BGGB4 for FFW, BGGB13 x BGGBI for FNFW, and BGGB12 BGGB6 for HF/W For 5-parenta Galle, the significant SCA effects were BGGB18 BGCB16 for DRMF, BOOB19 BGGB17 for DRFF, BOGB19 BOGB17 for FL, BOGB17 BGGB16 for FV, BOGB20BGGB19 for FFT and BGGB18 BOOB16 for SWF. For the traits HL, EL, LPL2, LBL2, LP, LBL, LBB, IL, FFN, OL, OW, SL, FD, FFW, SP and FP had no significant cresses. The best areas was BOOB 20x BOGB17 for most of the anatomical traits A high frequency of sprouting from shoot tips (90%) and nodal explants (93%) and shoot differentiation was observed in the primary culture of Momordica charantia L on MS hal medium supplemented with various concentrations and combinations of benzyl adenine (BAP), a-naphthalene acetic acid (NAA), indole-3-butyric acid (IBA) and gibberellic acid (GA). The best response in terms of single shoot formation was observed on MS medium with 2 mg1 BAP + 0.2 mgl NAA and multiple shoots were obtained by culture of shoot tips and nodal explants on MS with BAP (2 mg/l)+ IBA (0.2 ng)+GAs (0.3 mg). This combination was suitable for regeneration and growth of shoots. About 99% rating was obtained from shoots cultured on MS supplemented with IBA (0.6 mg/l). Pls with well developed roots were transferred to soil with survival frequency of 70% The growth of meristem was observed on semisolid MS media when the culture media were supplemented with 0.05mg/1 KIN +0.1 mg/l GA, After three weeks, meristems were transferred to MS media supplemented with BA, KIN, IBA, NAA and IAA singly or in combination for shoot elongation and root initiation. Among the different treatments, MS + 1.0 mg/1 BA + 0.1 mg IBA + 0.3 mg/1 GA, showed good responses for shoot initiation with elongation. On the other hand good rooting was observed when 0.5 mg IBA and 0.1mg/ NAA were used in MS semisolid medium. Ten works old in vitro plantlets were successfully planted in soll through gradual acclimatization. Immature embryos were excised from seeds of two genotypes (GBBG 6 and GBBG 14) of bitter gourd and cultured in invariably underwent callasing followed by multiple shoot formation, but in some cases single shoot was observed MS medium supplemented with 1 μM BAP + 0.01 M1 NAA was found to be suitable for adventitious multiple shoots and root regeneration. Addition of 40-100 mg/l casein hydrolysate (CH) improved shoot and root regeneration efficiency. For single shoot and root formation, MS medium with 0.3521 M1 GA+ 100 ml CH was very effective Aspangine and L-glutamine had no role in to tissue differentiation of immature embryo culture. High frequency of direct organogenesis from immature leaflet explants has been achieved in biter gourd. Here shoot buds were found to develop from leaflet on MS medium containing 3.0mg/l BAP and 0.5mgl IAA within 7-8 weeks. Proliferation of regenerated shoot buds, formation of shoots and their elongation took place in the same medium. Regenerated shoots formed healthy roots when they were further grown on MS median supplemented with 0.5mg IBA. Mutiple shoots from the immature cotyledenory nodes of two genotypes of bitter gourd were induced in MS media with GA, IAA, IBA, NAA, KIN and BAP. The shoots continued to increase in number for 5-6 weeks. On the medium, BAP 2 mg in combination with 0.1 mgl IAA + 2 mg/l GA, was the most effective medium for adventitious shoot proliferation from immature cotyledonary node. BGGB1 and BOGB14 genotypes produced over 84% and 80% shoots, respectively on this medium in 6 from the time at inoculation of primary explants. BOGBI was found to be better in rape than B00814 in most of the treatments. Somatic embryogenesis and organogenesis with subsequent plantlets regeneration were achieved in callus cultures derived from immature cotyledon of bitter gourd using MS semisolid besal medium supplemented with different concentrations and combinations of cytokinin (BAP and KIN) and on (IAA and IBA). Among the different combinations, the media supplied with BAP and IAA were more effective in the induction and proliferation of callus then KIN+ IBA or KIN+ IAA or BAP+ IBA. The highest numbers of somatic embryos were obtained in media fortified with IAA (1.0 mg/l) and BAP (1.0 mg). BAP (2.5 mg) in combination with 0.2 mg/l IAA was found table for shoot organogenesis. Rapid and reproducible transformation system for bitter gourd using Agro bacterium fciens mediated gene delivery was developed. Inmature cotyledonary node and immature cotyledon from green immature fruits that have been allowed to imbibe were used as explant, and regeneration we achieved vie organogenesis and embryogenesis, respectively, Cotyledonary node and cotyledon were co-cultured in MS media with 2.0 mgl BAP +0.1 mg IAA +2.0 mg/l GA and 2.5 mg BAP 0.2 mgl IAA, respectively. Histochemical GUS assay of the explants, exsand shoots (newly formed) released that they were GUS positive. Of the explants so the studied cotyledon was found to be more susceptible to Agrobactreium (LBA4404, pB1121) and infection moreover, successful regeneration was also observed. It was the first step to produce transgenic bitter gourd using this gene delivery method and it will also provide a basis for further research to generate transgenic bitter gourd plants.