Koike Y, Hoshino Y, Mii M, Nakano M
Faculty of Agriculture, Niigata University, 2-8050 Ikarashi, 950-2181 Niigata, Japan.
Plant Cell Rep. 2003 Jun;21(10):981-7. doi: 10.1007/s00299-003-0608-7. Epub 2003 Apr 2.
Genetic transformation was carried out with wild-type strains of Agrobacterium rhizogenes for introducing a dwarf trait into the Scrophulariaceous ornamental plant, angelonia (Angelonia salicariifolia). Leaf segments of two angelonia genotypes (Ang.1 and Ang.2) were co-cultivated with mikimopine-type strains of A. rhizogenes. Adventitious roots that showed vigorous growth and increased lateral branching when cultured on half-strength Murashige and Skoog's (MS) basal salts medium lacking plant growth regulators (PGRs) after co-cultivation were selected as putatively transformed lines. All of these selected lines produced mikimopine. Adventitious shoots were efficiently induced from putatively transformed root segments on half-strength MS basal salts medium containing 1 mg l(-1) benzyladenine (BA) under continuous illumination (24-h photoperiod), and the shoots easily rooted following their transfer to half-strength MS basal salts medium lacking PGRs. The transgenic nature of regenerated plants was confirmed by Southern hybridization. Transformed plants frequently died during their acclimatization, and acclimatized plants of eight transformed lines grew very slowly for 1-5 months after transplantation to the greenhouse. Plants of two transformed lines of Ang.2 flowered 4-6 months after transplantation. These transformed plants exhibited phenotypic alterations such as dwarfness and smaller leaves. There were no apparent alterations observed in the number, shape, and size of the flowers. Pollen fertility of the transformed plants was 60-80% based on aceto-carmine staining. These results indicate the possibility of applying A. rhizogenes-mediated transformation for introducing a dwarf trait into angelonia.
利用发根农杆菌野生型菌株进行遗传转化,将矮化性状导入玄参科观赏植物香彩雀(Angelonia salicariifolia)。将两种香彩雀基因型(Ang.1和Ang.2)的叶片切段与发根农杆菌的甘露碱型菌株共培养。共培养后,在不含植物生长调节剂(PGR)的1/2强度Murashige和Skoog(MS)基础盐培养基上培养时,选择生长旺盛且侧枝增加的不定根作为推定转化株系。所有这些选定的株系都产生了甘露碱。在连续光照(24小时光周期)下,在含有1 mg l(-1)苄基腺嘌呤(BA)的1/2强度MS基础盐培养基上,从推定转化的根段高效诱导出不定芽,并且这些芽在转移到不含PGR的1/2强度MS基础盐培养基后很容易生根。通过Southern杂交证实了再生植株的转基因性质。转化植株在驯化过程中经常死亡,8个转化株系的驯化植株在移植到温室后的1 - 5个月内生长非常缓慢。Ang.2的两个转化株系的植株在移植后4 - 6个月开花。这些转化植株表现出矮化和叶片变小等表型改变。花的数量、形状和大小没有明显变化。基于醋酸洋红染色,转化植株的花粉育性为60 - 80%。这些结果表明利用发根农杆菌介导的转化将矮化性状导入香彩雀的可能性。