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Determination of Some Alkaloids in Catharanthus roseus L. Callus In Vitro

تقدير بعض القلويدات في كالس نبات عين البزون .Catharanthus roseus L خارج الجسم الحي

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 Publication date 2018
  fields Groves
and research's language is العربية
 Created by Shamra Editor




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This investigation was carried out at Tissue Culture Laboratory, Department of Horticulture, University of Mosul, Iraq during 2012-2014) on Catharanthus roseus Variety; Nirvana Pink Blush, which is one of the important medicinal plant, containing a group of alkaloids. Parts of leaves, nodes, and internodes were cultured on MS medium supplemented with NAA for callus initiation, data were taken after 28-day results refer that highest weight of callus; 0.612g, 0.639g ,0.835g were obtained from cultivated parts of leaves, nodes and internodes on MS medium supplemented with 1.0mg/l NAA respectively. Callus was initiated and cultured using parts of leaves, nodes and internodes on MS medium supplemented with BA and 1.0 mg/l of NAA, after 50 days.


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Research summary
أجريت هذه الدراسة في مختبر زراعة الأنسجة النباتية بجامعة الموصل بالعراق خلال الفترة من 2012 إلى 2014 على نبات عين البزون (Catharanthus roseus) صنف Nirvana Pink Blush، وهو نبات طبي يحتوي على مجموعة من القلويدات. تم زراعة أجزاء من الورقة والعقد والسلاميات على وسط موراشيج وسكوغ (MS) المجهز بنفثالين حمض الخليك (NAA) لاستحداث الكالس. أظهرت النتائج أن أعلى وزن للكالس تم الحصول عليه من زراعة أجزاء الورقة والعقد والسلاميات على التوالي عند تركيز 1.0 مغ/ل NAA. تم تحليل مركبات القلويدات باستخدام جهاز HPLC، حيث تم تحديد مركبات الفينبلاستين والفينكريستين والفندولين والأجماليسين. أظهرت النتائج أن أعلى قيمة امتصاص لمركب الفينبلاستين كانت عند زمن احتباس 12.686 دقيقة من مستخلص كالس الورقة، وأعلى قيمة امتصاص لمركب الفينكريستين كانت عند زمن احتباس 17.980 دقيقة من مستخلص كالس السلامية. بينما كانت أعلى قيمة امتصاص لمركب الفندولين عند زمن احتباس 21.113 دقيقة من مستخلص كالس الورقة، وأعلى قيمة امتصاص لمركب الأجماليسين عند زمن احتباس 32.510 دقيقة من مستخلص كالس العقدة.
Critical review
دراسة نقدية: تعتبر هذه الدراسة مهمة في مجال زراعة الأنسجة النباتية وتحديد القلويدات في نبات عين البزون. ومع ذلك، يمكن تحسين الدراسة من خلال تضمين مزيد من التفاصيل حول الظروف البيئية التي تم فيها إجراء التجارب، مثل درجة الحرارة والرطوبة، وكذلك تأثير هذه العوامل على نمو الكالس وإنتاج القلويدات. بالإضافة إلى ذلك، يمكن توسيع الدراسة لتشمل تأثير أنواع أخرى من منظمات النمو أو الظروف البيئية المختلفة على إنتاج القلويدات. كما أن استخدام تقنيات تحليلية أخرى بجانب HPLC يمكن أن يضيف قيمة إضافية للدراسة.
Questions related to the research
  1. ما هو الهدف الرئيسي من الدراسة؟

    الهدف الرئيسي من الدراسة هو استحداث وتتميّة الكالس من زراعة أجزاء من الأوراق والعقد والسلاميات لنبات عين البزون وتقدير بعض القلويدات في الكالس الناتج باستخدام جهاز HPLC.

  2. ما هي أعلى قيمة امتصاص لمركب الفينبلاستين وأين تم الحصول عليها؟

    أعلى قيمة امتصاص لمركب الفينبلاستين كانت عند زمن احتباس 12.686 دقيقة وتم الحصول عليها من مستخلص كالس الورقة عند المعاملة بـ 0.3 مغ/ل BA مع 1.0 مغ/ل NAA.

  3. ما هي الظروف البيئية التي تم فيها زراعة الأجزاء النباتية؟

    تم زراعة الأجزاء النباتية في زجاجات حجم 200 سم³ من الوسط الغذائي عند pH 5.8، وتم تعقيم الوسط الغذائي عند درجة حرارة 121 مْ وتحت ضغط 1.04 كؤ/سم² لمدة 20 دقيقة. بعد الزراعة، نُقلت الزرعات إلى غرفة النمو تحت شدة إضاءة 3000 لوكس وبتعاقب يومي 16 ساعة ضوء و8 ساعات ظلام بدرجة حرارة 25 ± 2 م°.

  4. ما هي التوصيات التي قدمتها الدراسة؟

    توصي الدراسة بدراسة تأثير بعض الأحماض الأمينية أو التعريض لأشعة الليزر أو المجال المغناطيسي في نمو الكالس وكمية القلويدات الناتجة.


References used
Azimi, A. A.; B.D. Hashemloian; H. Ebrahimzadeh; and A. Majd (2008). High in vitro production of anti-canceric indole alkaloids from periwinkle (Catharanthus roseus) tissue culture. African J. of Biotechnology. 7(16): 2834-2839
Budhiraja, R.P. (2004). Separation chemistry new age international (P) Ltd., Publishers, PP. 424
Gilman, E.F.; and T. Howe. (1999). Catharanthus roseus. University of Florida - Cooperative Extension Service. Institute of Food and Agricultural Science. Fact Sheet FPS-112
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Experiments were conducted to study effect of radiation and salt levels on plant regeneration from callus for two cultivars of potato i.e. Riviera and Burren, under in vitro condition. Results showed that the efficiency of radiation in induced reg eneration from callus in Riviera cultivar at salt level of (10 dS m-1) and Burren cultivar at salt levels (8, 12 dS m-1). For the purpose of making sure inheritance of salinity tolerant, mutant clones (plants induced from salt tolerant calli, which planted at salt levels of 8, 10, 12 dS m-1 and plants induced from non salt tolerant calli, which planted at salt level of 6 dS m-1) and their parental cultivars (Riviera and Burren) by exposing to salt stress conditions and comparing them with control (6 dS m-1). Results revealed that the lowest percentage of reduction in plant height, number of nods per plant and tuberization were observed in salt tolerant mutants. Plant height and number of nodes/plant can be considered as selective morphological markers for in vitro salt tolerance .
The Madagascar periwinkle (Catharanthus roseus) produces numerous indole alkaloids, several of which have an important pharmaceutical uses such as ajmalicine. The relationship between hypoxia and ajmalicine production in a cell medium culture of C atharanthus roseus were investigated during the cycle of cell culture, as well as the growth in fresh and dry matter. The results show that the lack of oxygenation in C20D cells provokes a very strong inhibition in accumulation of the alkaloids and of other possible substances. This can be explained by the absence of some downstream enzymes involved in the biosynthesis chain of alkaloids or by the blockage of the upstream terpenes to the production stage. Reoxygenation did not restore hypoxia effect on the alkaloid accumulation. Moreover, the present study showed that the addition of the loganin in the 4th day, in the cell culture medium subjected to hypoxia restored the alkaloid production. Also, the results show that the BAP increases the ajmalicine production. The effect of hypoxia was not shown in the presence of cytokinine. The BAP can without doubt decrease the effects of the hypoxia and increases the ajmalicine production.
Stages of initial callus and embryiods of date palm Phoenix dactylifera L. were histological and chronological studied begging from the ٢nd weeks of culturing segment of shoot tip to ten months in MS medium containing ١٠٠mg/l ٢،٤-D with activated charcoal in the dark. The study reveled that the active meristematic cells were unequally distributed in the explant and concentrated in the shoot apical dome and leaf margin. These cells are the bases from which initial callus started its growth. Somatic proembryo appeared for the first time as a meristematic cells groups separated from each other by relatively thick walls after five months of culturing in continuos culture in the same medium .In progress months (٦-١٠) the small simple proembryos continued into advanced multicell embryos and underwent polarization. Also the embryos were observed with attendance for division and separation into smaller or budded to produce numerous other.
This research was conducted at the Laboratory of Tissue Culture, Faculty of Agriculture, Tishreen University. Because of the importance of the food and economic of chickpea, two Winter chickpea cultivars (Ghab 4, Ghab 5) and two Vernal chickpea culti vars were used (ILC263, ILC1929) and planted in pots until flowering, so the floral buds were picked before opened and washed with distilled water and alcohol three times. Anthers were separated and treated at preliminary temperatures (4 Cْ for 48 hours, and 35 Cْ for 12 hours), then it washed with a solution of hypochlorite of sodium( Naocl 2%) for a period of 15 minute, and washed with distilled sterilized water ,then where planted on the environment Murashige and Skoog ( MS) equipped with 1, 3 and 5 mg/L of auxin 2,4, D, as well as 0.1, 0.2 and 0.3 mg /L of Cytokinin Benzyl Amino Purine( BAP) individually and with interaction between them and incubated under conditions of 27 Cْ and 75% of the humidity and the intensity of 1500 Lux of light for 16 hours. The main objective of research was to study the effect of both preliminary heating treatment , quality and concentration of used hormone on producing callus from used chickpea cultivars anthers. Results showed difference in cultivars response in the treatment of both temperature and single hormone, so Ghab 5 cultivar was the most responsive to the formation of callus, while the results indicated that the highest percentage of the formation of callus was 40% when treated at 4 Cْ of Ghab 4 and Ghab 5 cultivars, while ILC263 cultivar showed less response in all the individual treatments for growth regulators. As well as results showed that the significant and catalyst role for using of hormones together with heat treatment, where each of Ghab 5, Ghab 4 and ILC1929 cultivars were the superiority, by the arrival of the percentage of 80% and 60%, respectively.
The environmental factors may have an effect on the contents of the active materials in plants. Salinity is one of these factors that can have an influence through the calcium ion which competes with the Na+, which in turn, affects the manufacture of alkaloids present in the Datura stramunium.
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