The present study describes plant regeneration via somatic embryogenesis from cotyledonary node callus. Callus cultures were derived from cotyledon node segment excised from Trigonella foenum-graecum L. axenic seedlings grown in agar solidified Murashing and Skoog's(MS) medium supplemented with Benzyl adenine BA at 2.0 mg l-1 and Naphthalene acetic acid NAA at 1.0 mg l-1. These cultures were stimulated to form somatic embryoids at 32% frequency of in agar-solidified MS medium containing NAA at 0.5mg l-1 in the presence of 15% v/v coconut water. Light microscopy examination of sections of heart embryos stage proved the presence of vascular system. Transfer of other developmental stages of embryos to the same induction medium led to the development of secondary somatic embryos and embryoids into plantlets in agar-solidified MS medium lacking hormones. They were readily rooted in MSO and successfully transferred to soil.
Inoculation of leaves and stems explants excised from field-grown Trigonella foenum-graecum L. with engineered Agrobacterium rhizogenes 1601 inoculum of optical density 1.90, 2.06 and 1.96 led to the formation of hairy roots on these explants. The highest percent of infection with inoculum of O.D 1.90 was 20% in leaves, and was 53.3% in stems using inoculum of O.D 2.06. Infection percent was 50% in hypocotyl compared with non-inoculated explants. The results showed that inoculation of seedlings lacking roots were slow in hairy root formation and take long time compared with their formation in intact seedlings inoculated with the same inocula. Results of paper electrophoresis of these root proved the incidence of their genetic transformation. Moreover, these transformed roots grow happily in liquid WP medium with an average fresh weight 5.9g after four weeks of culture, whereas fresh weight of normal roots was 1.02g under the same conditions.
نفذت هذه التجربة لفصل مركبات الستيرويدات (Steriods), احد نواتج الايض الثانوي لنبات كرز الارض Physalis angulata L.) ) من اوراق النبات عند مرحلة الازهار ومن الكالس المستحث من السيقان تحت الفلقية للبادرات المعقمة ومن الوسط الغذائي للمزرعة المستمرة المغلقة بعمر 7 أو 14 أو21 يوماً والخلايا المحصودة من المزارع الكمية, اذ اظهرت بيانات الكشف عن مستويات مركبات Physalin A و Physalin B في هذه المزارع, وجودها بدلالة قراءات كروموتوكرافيا السائل عالي الكفاءة HPLC بمقارنتها مع العينات القياسية. بلغ تركيز Physalin A و Physalin B في كل من الاوراق 24.36 و34.18 مايكروغرام.مل-1 على التوالي, وفي مزرعة الكالس بعمر 30 يوم بلغ تركيز 287.28 و 238.47 مايكروغرام.مل-1 على التوالي, وفي عينات الوسط الغذائي السائل للمزارع المستمرة المغلقة للمراحل العمرية 7 أو 14 أو21 يوم بلغ تركيز Physalin A 97.67 أو 137.88 أو 85.79 ميكروغرام.مل-1 على التوالي, و لـ Physalin B بلغت 103.36 أو194.19 أو40.96 مايكروغرام.مل-1 على التوالي, وبلغ اقل تركيز لـ Physalin A و Physalin B في الخلايا المحصودة من المزارع الكمية 10.11 و 15.59 مايكروغرام.مل-1 على التوالي. وهذه البيانات تشير الى حقيقة امكانية انتاج مركبات Physalin A و Physalin B في مزارع الانسجة كمصدر ثابت ودائم للحصول على نواتج االايض الثانوي وبديلا مناسبا عند النباتات الحقلية.
Thediosgenin compound was separated, one of the secondary metabolic product in Trigonella foenum-graecum L. plants. Hairy roots cultures transformed by Agrobacterium rhizogenes R1601, and their spontaneously formed callus contained "diosgenin". High performance liquid chromatography HPLC measurements proved the presence of diosgenin in these cultures that approach 158.3 and 156.5% respectively. It present in leaves of plants produced from seeds at 161.8%. Generally, these data pointed out that transformed tissues are often preferred as a continuous source of diosgenin and perhaps other secondary metabolites.
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