摘要
葡萄扦插技术不能获得大量的脱毒苗，传统的育种技术很难获得抗病葡萄品种，组织培养技术对葡萄无病毒株系建立、转基因葡萄育种研究提供了技术支持。而葡萄转基因工程目前发展的瓶颈是葡萄再生频率低，转化率低。本试验对酿酒葡萄赤霞珠、黑比诺和霞多丽离体茎尖、叶片和叶柄再生进行了研究，并对影响再生的基因型、植物激素和培养基组分等因素进行了探讨。主要取得了以下研究结果：
1．通过对赤霞珠、黑比诺和霞多丽离体茎尖繁殖体系建立的研究，获得茎尖增殖的最佳培养基配方。赤霞珠茎尖增殖培养基为MS+BA1.5mg/L+NAA0.02mg/L；黑比诺和霞多丽茎尖增殖培养基为MS+BA1.0mg/L+NAA0.02mg/L。同时，获得了赤霞珠、黑比诺和霞多丽茎尖增殖芽生根培养基配方。赤霞珠茎尖增殖芽生根培养基为1/2MS+IBA1.0mg/L；黑比诺和霞多丽茎尖增殖芽生根培养基为1/2MS+IBA0.5mg/L。
2．通过对赤霞珠、黑比诺和霞多丽离体叶片、叶柄再生体系建立的研究，获得了赤霞珠、黑比诺和霞多丽离体叶片再生不定芽培养基配方。赤霞珠、霞多丽叶片再生培养基均为MS+TDZ4.0mg/L时，再生效果最好，其再生诱导率分别为18.1％、10.0％。黑比诺在MS+TDZ6.0mg/L+NAA0.05mg/L时，再生稳定，诱导率为6.7％。同时获得了赤霞珠叶柄离体再生不定芽的培养基为MS+TDZ4.0mg/L+NAA0.01mg/L，其再生诱导率为28.6％。获得了赤霞珠离体叶片不定芽生根苗的培养基配方，为1/2MS+IBA1.0mg/L。
3．试验结果表明，细胞分裂素BA对赤霞珠、黑比诺离体叶片愈伤组织诱导和不定芽再生基本无作用，细胞分裂素KT对赤霞珠、霞多丽离体叶片愈伤组织诱导和不定芽再生基本无作用。同时发现增加细胞分裂素的浓度和种类并不能提高离体叶片再生诱导率。外植体材料的选择对离体再生有较大的影响，带叶柄砧的叶片比不带叶柄砧的叶片诱导率要高。
Grape cuttage can not gain the virus-free plant, and it is very difficult to get the resisting diseases variety in traditional hybridization way, while tissue culture technology can finish the requirement of virus-free plant establishment and transgenic breeding, but the hinder of grape genetic engineering is that regeneration rate and transformation rate is too low.
Good result was obtained by studying the tips, leaves, and petioles regeneration of Carbernet Sauvignon, Pinot Noir and Chardonnay , and studying the influence of the genotype, plant hormone, and cultural medium composition.
1. By studying on the Carbernet Sauvignon, Pinot Noir and Chardonnay's shoot tips propagation, we obtained the best propagation culture medium ( Carbernet Sauvignon:MS+BAl. 5mg/L+NAA0. 02mg/L; Pinot Noir and Chardonnay: MS+BA1. 0mg/L+NAA0. 02mg/L) . By studying on the Carbernet Sauvignon, Pinot Noir and Chardonnay's propagation shoots rooting , we obtained the best rooting culture medium(Carbernet Sauvignon: 1/2MS+IBA1. 0mg/L ; Pinot Noir and Chardonnay: 1/2MS+IBA0. 5mg/L).
2. By studying on the Carbernet Sauvignon, Pinot Noir and Chardonnay's leaves , petioles regeneratoion , we have obtained the leaves regeneration culture medium of Carbernet Sauvignon(MS+TDZ4.0mg/L), Chardonnay(MS+TDZ4.0mg/L), Pinot Noir (MS+TDZ6. 0mg/L+NAA0. 05mg/L) and their regeneration rates is respectively 18.1%, 6. 7%, 10.0%. We also obtained Carbernet Sauvignon petioles regeneration culture medium(MS+TDZ4.0mg/L+NAA0.01mg/L) and it's regeneration rate is 28.6%, and obtained the rooting medium of Carbernet leaves regeneration buds (1/2MS+IBA1.0mg/L).
3. The research indicates that the cytokinin BA have no effect on the leaves regeneration of Carbernet Sauvignon, Pinot Noir , and the KT have no effect on the leaves regeneration of Carbernet Sauvignon, Chardonnay. Increasing the cytokinin concentration and kinds do not raise the regeneration rate of the leaves in vitro, and also find that selection of explant is very important to the explant regeneration, and regeneration rate of leaves with petiole stock is higher than the leaves without petiole stock.
引文
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