葡萄組織培養之研究　環境因子對葡萄癒傷組識生長及分化的影響

Studies on Tissue Culture of Grape (Vitis viniferal L.) Influence of Environmental Factors on the Growth and Differentiation of Grape Callus.

賴本智(Pen-Chih Lai) ； 易希道(Shi-Tao Yie)

《中華農業研究》 29卷2期 (1980/06) Pp. 157-165

https://doi.org/10.29951/JARC.198006.0010

摘要

巨峰葡萄（Vitis viniferal L. CVS. Sentemal X石原早生）植株的莖段，葉片及果肉培養於含NAA 0.1mg/l、Kinetin 0.2mg/l及15%(w/v) coconut milk.的Gamborg和Eveleigh基本培養基中可誘導形成癒傷組織，經上述培養基繼代培養得多量癒傷組織，以進行各種環境因子對其生長及分化影響的測定。經研究顯示，莖段癒傷組織生長最適的PH值爲5.6。光線的強弱能影響癒傷組織的生長，果肉癒傷組織適合生長於750到3,000lux之間的光強，光強超過4,500lux或在黑暗下培養均不利於癒傷組織的生長。在各種光質中以藍光最適於果肉癒傷組織的生長，黃光、紅光次之，綠光較差，固體培養基培養之葉片與莖段癒傷組織均可形成烏巢狀的維管束環節(nest-like Vascular nodule)，其內部具有許多木質化的細胞或管形細胞，外部則具有許多類似形成層細胞(Cambium-like cells)的環狀結構，由此類形成層結構，可再進一步分化形成根。而葉片癒傷組織外部的細胞也可分化形成芽原體(shoot primordial)之構造。果肉癒傷組織在任何培養情形下，均無分化的細胞。液體震盪培養下的葉片和莖段癒傷組織其細胞僅可分化形成似根之多極性(poly-arch)維管組織型(Vascular pattern)，無再進一步分化的現象。而果肉癒傷組織則均呈游離的細胞或細胞羣，沒有癒傷組織塊的形成，也無分化的現象。液體震盪培養雖能誘發葉片及莖段癒傷組織的特化，唯較固體培養者差。

並列摘要

Callus from grape stem, leaf and fruit sections were induced in modified Gamborg and Eveleigh (1968) medium containing 0.1 mg/l of NAA, 0.2 mg/l of kinetin and 15% (w/v) of coconut milk. They were then subcultultured for testing the influence of medium pH and light factors on the callus growth.
Optimum pH of medium for the growth of stem callus was found to be 5.6 followed by 5.2. Values above 6.4 or below 4.8 were not suitable for weight increases of callus in culture.
Fruit callus showed maximum growth when grown under blue light. Vigorous growth was also found under red and yellow light. However, growth was limited when callus were subjected to green light or darkness treatment. A light intensity of 3,000 lux was observed to give the hightest growth rate of fruit callus followed by 1,500 and 750 lux. Higher (4,500 lux) or lower (250 lux) light irradiation was less effective in increasing both the fresh and dry weight of callus.
Histological studies showed that when grown on solid medium, callus cells originated from stem sections could differentiate into nest-like vacsular bundles which contained many lignified cells and tracheid elements in the interior. Some nodulous structures could further differentiate into roots. Shoot primordia from the peripheral layer of leaf callus were also recorded. Fruit callus showed no ability of differentiation.
In liquid culture, the leaf and stem callus were able to form poly-arch vascular bundles but showed no sign of organ differentiation. Fruit callus cultured in liquid resulted in many free cells or cell clonies without any specific differentiation. It was concluded that the regeneration ability was much higher when grown on solid than in liquid medium.