การขยายพันธ์ุต้นกะเพราหินปูน (Coleus albicalyx (Suddee) Suddee) พืชหายากของประเทศไทยในหลอดทดลอง
In vitro Propagation of Coleus albicalyx (Suddee) Suddee, a Rare Plant of Thailand
Keywords:
พืชถิ่นเดียว , การเพาะเลี้ยงเนื้อเยื่อพืช , เบนซิลอะดีนิน , อาการฉ่ำน้ำ , endemic plant , plant tissue culture , N6-benzyladenine , hyperhydricityAbstract
งานวิจัยนี้เสนอวิธีการขยายพันธุ์ในหลอดทดลองของต้นกะเพราหินปูน (Coleus albicalyx (Suddee) Suddee) ซึ่งเป็นพืชที่พบเฉพาะในประเทศไทยเท่านั้นและมีสถานะเป็นพืชหายากของประเทศไทย โดยนำเมล็ดมาผ่านกรรมวิธีการฟอกฆ่าเชื้อผิวและเพาะบนอาหารสังเคราะห์สูตร Murashige และSkoog (MS) หลังจากได้ต้นกล้าอายุ 8 สัปดาห์ที่มีความสูง 4 เซนติเมตรจึงนำต้นกล้าดังกล่าวมาตัดเอาส่วนลำต้นที่มีข้อแรกนับจากโคนลำต้นให้มีขนาด 1.5 เซนติเมตร แล้วนำไปเลี้ยงบนอาหารสังเคราะห์สูตร MS ที่มีเบนซิลอะดีนิน (N6-benzyladenine: BA) เข้มข้น 0 1 2 3 และ 4 มิลลิกรัม/ลิตร หลังจากเลี้ยงชิ้นพืชนาน 6 สัปดาห์ พบว่าอาหารสังเคราะห์สูตร MS ที่มี BA เข้มข้น 1 มิลลิกรัม/ลิตร เป็นสูตรอาหารที่เหมาะสมสำหรับการขยายพันธุ์ต้นกะเพราหินปูน เนื่องจากเนื้อเยื่อที่เลี้ยงบนอาหารสูตรนี้สามารถรอดชีวิตและชักนำให้เกิดยอดใหม่ได้ทุกชิ้นพืช ทั้งยังให้จำนวน (4.70 + 0.15 ยอด/ชิ้นพืช) และความสูงของยอดใหม่ (3.12 + 0.40 เซนติเมตร) ที่สูงที่สุดอีกด้วย นอกจากนี้ยอดใหม่ที่เกิดขึ้นจากสูตรอาหารข้างต้นยังไม่พบลักษณะฉ่ำน้ำ เมื่อนำยอดใหม่ที่เกิดขึ้นมาตัดแยกเป็นยอดเดี่ยวและนำไปเลี้ยงต่อบนอาหารสังเคราะห์สูตร MS อีก 6 สัปดาห์ พบว่ายอดพัฒนาเป็นต้นพืชที่สมบูรณ์ โดยมีการยืดยาวของลำต้นสูงขึ้นและพบการเจริญของรากเป็นจำนวนมาก ผลที่ได้จากการศึกษานี้สามารถใช้เป็นวิธีการอนุรักษ์พันธุ์ต้นกะเพราหินปูนนอกถิ่นอาศัยซึ่งเป็นการช่วยรักษาความหลากหลายทางชีวภาพของประเทศไทย และอาจใช้เป็นเครื่องมือในการผลิตพืชสมุนไพรชนิดนี้ให้ได้จำนวนมากเพื่อใช้เป็นวัตถุดิบในการศึกษาด้านพฤกษเคมีต่อไป This research presented an in vitro propagation method of Coleus albicalyx(Suddee) Suddee, anendemic plant and also classified as a rarespecies of Thailand. Seeds surface was sterilized before culturing on Murashige and Skoog (MS) medium. After the 8-week-old seedlings at 4 cm in height were achieved, the nodal segments contained the first node from the base of seedlings were excised to 1.5 cm in height. Then, these explants were cultured on MS medium augmented with 0, 1, 2, 3, and 4 mg/L of N6- benzyladenine (BA). The results at the 6th week of culture revealed that MS medium supplemented with 1 mg/L BA was the appropriate medium for multiplying new shoots C. albicalyx. All explants cultured on this medium could survive and produce new shoots. The best outcomesof new shootsnumber (4.70 + 0.15 shoots/explant) and height (3.12 + 0.40 cm) were also found from this treatment. Moreover, the regenerated shoots from this medium did not show hyperhydricity. These regeneratedshoots were excised into a single shoot and transferred to MS medium for further6 weeks. Shoots developed into complete plantlets by elongated their stem and generated profuse root growth. The outcome of this study can be used as a method for ex situconservation of C. albicalyx, which assists in the maintenance biodiversity of Thailand. Furthermore, it may be applied as a mass propagation tool of this medicinal plant to provide material for the phytochemical study.References
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