ABSTRACT

Kakrol (Momordica dioica Roxb) is an underutilized summer vegetable of cucurbitaceae family. It has high nutritional and medicinal value. It is a dioecious vegetable. The seed when planted segregate into 50:50 males:females. Since the economic part i.e fruit is present on female plant a ratio of 90:10 female:male plants is recommended for economic cultivation. But to go for organized ratio of planting there are no distinguishable morphological markers for identification of sex prior to flowering. Even there are no reliable biochemical markers for determination of sex. Hence, this particular study i.e use of molecular markers in determination of sex in kakrol using RAPD’s has been taken up.

Kakrol genomic DNA was isolated from the three different ages of leaves viz., terminal young leaves, medium matured leaves and fully matured leaves of vine. The young leaves were ideal for the isolation of quality DNA. Standardization of PCR depends on primer annealing to template DNA. Generally 25-50ng template sufficient for PCR. Annealing was effective at °C compared to 36°C. Eighty RAPD operon primers were screened for polymorphism for sex with DNA of one male and female. Of which seven primers viz., OPA7, OPA18, OPD3, OPD5, OPD12, OPD18 and OPF20 expressed polymorphism at single locus among females and males. Hence these primers were screened against number of males (5) and females (5) for confirmation of polymorphic band related to sex determination. These primers when screened against DNA of more number of male and female vines, presence of band which was thought to sex specific was not consistent. For eg: OPA7 in which female specific band was observed. When screened with DNA of more number of male and female vines, the female specific band appeared in DNA of males also. Similar results were observed with other sex polymorphic primers. Hence it is proposed to continue this study using more number of primers (in 100’s) and by screening more number of male and female plants. Also male and female vines may be developed by sibbing in controlled conditions with maximum level of uniformity except for sex (isogenic lines except for sex).