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利用RT-PCR技术研究了美洲黑杨(Populus deltoides)材性相关候选基因细胞分裂素结合蛋白(cytokininbin ding protein)基因PdCYTOB的表达谱,结果显示,在未成熟木质部、未成熟韧皮部和韧皮部中PdCYTOB基因具有较高水平的表达量。对导入反义PdCYTOB的山新杨(Populus davidiana×P.bolleana)植株进行Southern杂交和RT-PCR检测,证实反义PdCYTOB基因已整合到杨树基因组中并表达。对大田转基因株系及对照植株进行表型观察、组织切片和微纤丝角的测定,结果表明,转反义PdCYTOB基因植株的高度明显增加,木质部、韧皮部变宽,微纤丝角明显变小,初步表明转基因杨树在造纸性能上有所改良。这些研究结果对于阐明PdCYTOB在美洲黑杨木材形成中的分子作用机制具有重要的理论意义。
The expression profile of the cytokininbin ding protein gene PdCYTOB was studied by RT-PCR. The results showed that in the immature xylem, immature phloem and phloem PdCYTOB Genes have higher levels of expression. Southern hybridization and RT-PCR analysis of the Populus davidiana × P. bolleana plants introduced with antisense PdCYTOB confirmed that the antisense PdCYTOB gene was integrated into the poplar genome and expressed. The phenotypic observations, histological sections and microfibril angles of the transgenic and control plants were carried out. The results showed that the height of the antisense PdCYTOB gene was significantly increased, the xylem and phloem broadened, and the microfibril angle was significantly reduced , Preliminary showed that the transgenic poplar papermaking performance has been improved. These findings are of great theoretical significance for clarifying the molecular mechanism of PdCYTOB in the wood formation of Populus nigra.