Bruce W, Desbons P, Crasta O, Folkerts O
Pioneer Hi-Bred Intl., Inc., 7300 NW 62nd Avenue, Johnston, IA 50131-1004, USA.
J Exp Bot. 2001 Mar;52(Spec Issue):459-68. doi: 10.1093/jexbot/52.suppl_1.459.
To assist breeding for increased resistance to root lodging in maize, an attempt was made to identify genes that are associated with root lodging by profiling mRNA expression from two inbreds with contrasting root-related traits. These two inbreds were derived from a common F(2) pool, selfed for several generations and showed 75% relatedness based on 106 genetic markers. Under field conditions, the two inbreds exhibited significant differences in root morphology and resistance to root lodging. Whole root tissue was collected at two developmental stages from inbred 100 and 101 grown in 2 years. RNA was isolated from both the V8 and V12 stages, a few weeks prior to flowering. The RNA samples from the 1997 growing season were analysed by GeneCalling analysis, an open-ended mRNA profiling method. From over 13 500 cDNA fragments detected from each of the V8- and V12-stage samples, 229 and 325 cDNA fragments, respectively, showed greater than 2-fold differences between the two inbred lines. A total of 69 cDNA fragments that showed 2-fold or greater differences for both inbred lines were observed at both developmental stages. The gene identity and expression differences of several cDNA fragments were determined and confirmed by RNA gel blot analysis. Two genes out of five identified were homologous to a cytochrome P450 and the impedance-induced protein, both showing high levels of expression in the roots of lodging resistant lines and low levels in the sensitive lines. These data provide the first clues of genes expressed in the roots during the formative stages of root development associated with root-lodging resistance.
为了辅助玉米抗根倒伏育种,通过对两个具有不同根系相关性状的自交系进行mRNA表达谱分析,试图鉴定与根倒伏相关的基因。这两个自交系源自一个共同的F(2)群体,经过多代自交,基于106个遗传标记显示出75%的亲缘关系。在田间条件下,这两个自交系在根系形态和抗根倒伏能力上表现出显著差异。在两年中,从自交系100和101的两个发育阶段采集了整个根系组织。在开花前几周,从V8和V12阶段分离RNA。对1997年生长季的RNA样本采用基因呼叫分析(一种开放式mRNA谱分析方法)进行分析。从V8期和V12期样本各自检测到的超过13500个cDNA片段中,两个自交系之间分别有229个和325个cDNA片段表现出2倍以上的差异。在两个发育阶段共观察到69个在两个自交系中都表现出2倍或更大差异的cDNA片段。通过RNA凝胶印迹分析确定并证实了几个cDNA片段的基因身份和表达差异。在鉴定出的五个基因中,有两个与细胞色素P450和阻抗诱导蛋白同源,它们在抗倒伏系的根中高表达,在敏感系中低表达。这些数据为根系发育形成阶段与抗根倒伏相关的根中表达的基因提供了首批线索。