Abstract
To regenerate adventitious shoots from the cotyledon proximal parts of Citrullus lanatus (Thunb.) Matsum. and Nakai ssp. mucosospermus (Fursa) oleaginous type, different concentrations of MS mineral elements, sucrose, 6-benzylaminopurine (BAP) and agar were tested. Shoot induction proved to depend on the interaction between levels of sucrose, BAP and MS mineral elements in the medium. The medium containing 3/2 strength of MS mineral elements, 35 g/l sucrose and 1 mg/l BAP solidified with 6 g/l agar allowed the production of numerous shoots without a callus phase. After 3 weeks of culture, 76.7% of the cotyledon proximal parts induced shoots with an average of 12.26 shoots per explant and a mean shoot length of 17.13 mm. The induced shoots were directly rooted and thus complete plants ready for acclimatization were obtained using a two steps procedure. Depending on the genotype, the shoot induction from cotyledon proximal parts ranged from 54% to 96%. Rooted plantlets were acclimatized and transferred to field, where they grew well, developed flowers and fruits like seeded plants. The assessment of the genetic stability of the in-vitro-regenerated plantlets by means of an Amplified Fragment Length Polymorphism (AFLP) analysis with the combination of 5 primers revealed no differences between regenerated plantlets and mother plants.
Highlights
Citrullus lanatus (Thunb.) Matsum. and Nakai ssp. mucosospermus (Fursa) oleaginous type and other species of the Cucurbitaceae family are grown mainly for their seeds which constitute an important component of the diet in many African (Nigeria, Benin, Ghana, Cameroun, Congo, Namibia, Sierra Leone and Côte d’Ivoire) and Asian countries [1,2,3,4]
The purpose of this study is to develop for the C. lanatus oleaginous type a two-phase micropropagation protocol that is fast, efficient and highly reproducible
Morphological characterization of the micropropagated plants was compared with the seedpropagated plants, using standard descriptors for cucurbits: eight characters (Table 2) were chosen among those published for Lagenaria siceraria [25]
Summary
Citrullus lanatus (Thunb.) Matsum. and Nakai ssp. mucosospermus (Fursa) oleaginous type and other species of the Cucurbitaceae family are grown mainly for their seeds which constitute an important component of the diet in many African (Nigeria, Benin, Ghana, Cameroun, Congo, Namibia, Sierra Leone and Côte d’Ivoire) and Asian countries [1,2,3,4]. Increasing the use and the production of these cucurbits can improve food security and diversify small farmer’s income [4] In this endeavor the development of micropropagation protocols will offer an easy and economic alternative for the rapid multiplication of elite improved cultivars and the conservation and distribution of genetic resources of Citrullus lanatus oleaginous type. Most of the protocols developed for the micropropagation of the Cucurbitaceae species are carried out in more than two phases [19,20], e.g., a bud induction phase with/or a multiplication phase, an elongation phase, and a rooting phase This approach is time consuming, expensive and sometimes yields a low rate of shoot induction. The genetic stability of the in vitro plants regenerated was assessed by an AFLP analysis, a technique proven to be a highly efficient tool for characterizing somaclonal variation [21,22]
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