石榴2种外植体再生方法在遗传转化研究中的优势比较

【目的】为了利用‘突尼斯软籽’石榴开展遗传转化研究,【方法】分别以叶片和茎段为外植体,探讨了不同灭菌时间和不同植物生长调节物质对2种外植体再生的影响,并对2种愈伤组织再生体系进行了比较。【结果】结果表明,茎段比叶片更容易灭菌,但叶片更容易长出绿色致密愈伤组织并分化更多的不定芽,而茎段分化的愈伤相对较为疏松,分化的不定芽也相对较少;来自茎段愈伤组织的不定芽比来自叶片的更容易长高;叶片比茎段再生体系建立所需时间要短;少量翠绿的叶片刀口处可以直接产生不定芽,可以直接利用叶片进行遗传转化的侵染,简化愈伤形成这一步骤,而茎段未发现此现象。在遗传转化试验中,茎段和叶片2种外植体愈伤组织抗生素敏感性试验结果均无显著性差异。【结论】研究结果认为,2者相比,叶片愈伤组织再生体系更适宜于石榴的遗传转化研究。

Comparison of advantages of two kinds of explant regeneration methods for studies on genetic transformation in pomegranate

【Objective】 The leaf and stem segments of ’Tunisia soft-seed’ pomegranate were used as the explants for genetic transformation research.【Method】 The effects of different sterilization time and different growth regulators on the in vitro leaf and stem segment regeneration were studied,and the advantages of two regeneration systems were compared.【Results】 The results showed that it was easier for stem segments to be sterilized than for the leaf,but the green and compact callus from leaves were more likely to be obtained and then more adventitious buds differentiated.The callus from stem segments were crisp,and the adventitious buds were less.The adventitious buds from the stem segment callus grew more quickly,but the time consuming of leaf regeneration system was shorter.The adventitious buds could be induced from a few emerald green leaves,which could be used for transformation,but the adventitious buds could not be induced directly from stem segments.There was not any significant difference between stem and leaf explants in callus sensitivity to antibiotics.【Conclusions】 As a result,the leaf callus regeneration system might be more appropriate for the genetic transformation research of pomegranate.