抗稻瘟病细胞突变体筛选技术的研究进展

稻瘟病是发生在世界各稻区最严重的真菌性病害,做好对该病的防治对水稻高产与稳产至关重要。随着生物技术的迅速发展,组织培养的离体筛选技术作为一种获得抗病突变体的高效手段而被广泛应用于水稻抗病育种研究。综述了抗稻瘟病细胞突变体筛选中愈伤组织的诱导、粗毒素的制备及毒素对材料抗性筛选等3个主要技术环节,并对抗病突变体在抗病育种中的应用提出问题及展望。     

金钗石斛类原球茎转化子选择方式的比较

为确立农杆菌介导法转化金钗石斛类原球茎的适宜的选择体系,利用液体纸基、固体培养基与液体滤纸桥3种培养方式,以0,20,40,50,60,80 mg/L质量浓度的潮霉素(Hyg)梯度试验,发现不同培养方式的选择效果有所差异。当使用液体纸基培养在50 mg/L Hyg的选择压下,20 d后可使材料死亡;使用固体培养基时,60 mg/L Hyg的选择压下,1个月以上才见效;在40 mg/L的选择压下,液体滤纸桥选择需要26~29 d。固体培养基选择需要较高的Hyg质量浓度且选择周期长,液体纸基选择可缩短时间,但容易造成转化细胞的生长抑制甚至死亡,比较而言液体滤纸桥是较好的选择方式。     

利用正交设计优化金钗石斛的转化体系

为优化金钗石斛的转化体系,以其类原球茎(PLBs)为材料,利用正交设计研究了影响农杆菌介导转化的因素.结果表明:PLBs预培养1 d、侵染的菌液OD600值1.2、乙酰丁香酮(AS)100μmol/L、共培养5 d时,每100mg鲜物质量材料上的GUS染色蓝点数最多;单因子试验表明AS适宜的浓度为50μmol/L;PCR结果显示得到了转化呈阳性的PLBs;初步表明此体系可用于金钗石斛的遗传转化研究.

Abstract：

In order to optimize the Agrobacterium-mediated transformation system for Dendrobium nobile Lindl, effects of preculture time, bacterial concentration, acetosyringon (AS) concentration and co-cultivation time on Agrobacterium-mediated transformation of protocorm like bodies (PLBs) of D. nobile Lindl were studied in an experiment with orthogonal design. The results showed that number of GUS staining blue spots per 100 mg flesh weight of PLBs was the greatest in the treatment of the combination of preculture for one day, 1.2 of bacterial concentration (OD600), 100 μmol/L of concentration of AS and co-culture for five days. Single factor experiment showed that the appropriate concentration of AS was 50 μmol/LPCR confirmed that positive transformed PLBs were obtained. It is tentatively concluded that this system can be used in the study of transformation of D. nobile Lindl.