RNAi沉默BcMF3基因对菜薹花粉发育的影响

园艺学报 ›› 2007, Vol. 34 ›› Issue (1): 125-130.

RNAi沉默BcMF3基因对菜薹花粉发育的影响

刘乐承^1,2 ; 向珣¹ ;曹家树^1*

(¹ 浙江大学蔬菜研究所, 杭州310029; ² 长江大学园艺园林学院, 湖北荆州434025)

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-02-25 发布日期:2007-02-25

Effects of Silenc ing BcMF3 by RNAi on Pollen Development of FloweringChinese Cabbage

LIU Le-cheng^1,2 ； XIANG Xun¹ ； CAO J ia-shu^1*

( ¹ Institute of Vegetable Science, Zhejiang University, Hangzhou 310029, China; ² College of Horticulture and Gardening, YangtzeUniversity, Jingzhou, Hubei 434025, China)

Received:1900-01-01 Revised:1900-01-01 Online:2007-02-25 Published:2007-02-25

摘要/Abstract

摘要： 根据从白菜(Brassica campestris ssp. chinensis var. communis) 核雄性不育两用系中分离到编码
果胶甲酯酶( PME) 的雄性不育相关基因BcMF3 cDNA序列设计引物, 从白菜花蕾cDNA中扩增出一短一
长两个片段, 分别反向和正向连接至双元载体pB I12 l中, 得到了RNAi (RNA interference) 植物表达载体
pB I2B3R, 并导入农杆菌LBA4404菌株中; 通过组织培养途径转化菜薹(B. campestris ssp. chinensis var.
parachinensis) , 分子检测证明B cM F3片段单拷贝整合到转基因菜薹的基因组中; 50%转基因菜薹植株的花
粉畸形, 花粉离体萌发率为32。3% , 出现畸形花粉植株的花药PME活性降低了13。5%。这一结果从转基
因植株后代的表型和酶活性上证明, 采用RNAi技术沉默BcMF3基因, 可能通过影响PME活性而引起转基
因菜薹植株部分花粉的败育, 从而证明BcMF3基因在普通白菜和菜薹等植物的花粉发育中起着重要作用。

关键词: 白菜, 菜薹, 花粉

Abstract: In an effort to study the molecular mechanism of genic male sterility in plants, BcMF3 gene
that encodes a pectin methylesterase ( PME) was isolated from the fertile B line of Chinese cabbage-pak-choi
(Brassica campestris ssp. chinensis var. communis, syn. B. rapa ssp. chinensis var. communis) previously.
In the present paper, primerswere designed based on the cDNA sequence of B cM F3, and a 458 bp fragment
and a 538 bp fragmentwere amp lified from the cDNA of flower buds of Chinese cabbage-pak-choi. These two
fragmentswere introduced into binary vector pB I121 in antisense orientation and sense orientation respectively,
and the generated RNA interference (RNAi) vector was then mobilized into Agrobacterium tum efaciens strain
LBA4404. The A. tum efaciens harboring the BcMF3 fragment was transformed to flowering Chinese cabbage
(B. campestris ssp. chinensis var. parachinensis) via tissue culture. The pollen grains from app roximately
50% of RNAi plants exhibited abnormal shapes, and only 32.3% of pollens from RNAi plants germinated nor
mally. The PME activity of the anther from RNAi plants with abnormal pollens decreased by 13.5%. These
results showed that functional interruption of BcMF3 by RNAi resulted in pollen abortion in flowering Chinese
cabbage, which p robably caused by the decrease of PME activity. The study suggests that the product of
BcMF3 gene plays an important role during pollen development in Cruciferae crop s such as flowering Chinese
cabbage and Chinese cabbage-pak-choi.

Key words: Chinese cabbage-pak-choi, Brassica campestris, Flowering Chinese cabbage, Pollen

刘乐承;;向珣;曹家树. RNAi沉默BcMF3基因对菜薹花粉发育的影响[J]. 园艺学报, 2007, 34(1): 125-130.

LIU Le-cheng;;XIANG Xun;CAO J ia-shu. Effects of Silenc ing BcMF3 by RNAi on Pollen Development of FloweringChinese Cabbage[J]. ACTA HORTICULTURAE SINICA, 2007, 34(1): 125-130.

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