摘要
菌核病严重威胁着我国长江中下游地区甘蓝型油菜的生产 ,几乎每年都会导致甘蓝型油菜的大面积减产。来自于烟草的葡聚糖酶 (Glucanase)基因 ,其编码产物能降解真菌的细胞壁结构成分 -葡聚糖。来自于烟草的植物蛋白AP2 4 则属于细胞程序性死亡 (PCD)的诱导蛋白 ,对多种真菌具明显抑制作用。前期已成功地将这两个广谱抗病基因导入甘蓝型油菜 ,并且通过表型鉴定 ,在T0 和T1代检测到了核盘菌抗性。本实验对转基因T2 代和T3代甘蓝型油菜进行分子鉴定和田间抗病性鉴定。经过PCR分子鉴定 ,从 14个转基因T3代甘蓝型油菜株系中筛选到 4个外源基因已经纯合的株系。对株系GA0 2 - 2的RT PCR鉴定则进一步证明外源广谱抗病基因在RNA水平上的大量表达。田间抗性鉴定主要利用苗期离体叶接种和成株期茎杆插签接种这两种方法。结果表明 ,T2 代转基因株系中有 5个株系表现为抗病基因初步纯合 ,5个株系表现为隐性纯合 ,12个株系处于分离当中 ,其中株系GA0 2 - 2的抗病性鉴定结果与RT PCR鉴定的结果吻合。 18个T3代转基因甘蓝型油菜株系中有 7个株系表现为抗性纯合 ,3个株系表现为隐性纯合 ,其余 8个株系基本上仍在分离中 ,其中株系GA0 2 - 2 - 1的抗病性鉴定结果与前面的分子鉴定以及上代的抗病性鉴定完全吻合。综合分子鉴?
Sclerotinia sclerotiorum is the most devastating disease of rapeseed (Brassica napus L ) in China especially in Yangtze River Area and leads to great yield loss every year. Glucanase and AP 24 , which come from tobacco both, have broad spectrum of antifungal activity. Our laboratory had transformed these two genes into Brassica napus and obtained Sclerotinia sclerotiorum resistance in T 0 and T 1 plants. This work took two different inoculation methods during two different periods to estimate the resistance to Sclerotinia sclerotiorum of T 2 and T 3 progenies. That is detached leaves inoculation and inoculation of stem by tooth sticks with clinging pathogene. The plants exhibited an increased resistance against the fungal attack compared with the nontransgenic plants during two inoculations. The candidate plants were also analyzed by PCR and RT PCR. Combined these methods, homozygous transgenic plants were selected.
出处
《分子植物育种》
CAS
CSCD
2003年第4期457-463,共7页
Molecular Plant Breeding
