OsMAX1a, OsMAX1e通过参与独角金内酯的合成调控水稻分蘖

doi:10．3969/j．issn．1001G7216．2015．03．001

摘要/Abstract

摘要：

独角金内酯是调控水稻分蘖的重要激素,在拟南芥中AtMAX1编码细胞色素P450蛋白,是独角金内酯合成途径的重要基因。通过同源序列比对,发现在水稻中有5个MAX1同源基因,包括OsMAX1a、OsMAX1b、OsMAX1c、OsMAX1d和OsMAX1e。分析5个OsMAX1基因的转录水平,发现OsMAX1a和OsMAX1e基因能够在水稻组织中稳定高效表达。对OsMAX1a和OsMAX1e的基因功能进行了验证,在拟南芥中过量表达OsMAX1a和OsMAX1e基因能够使max1突变体多分支表型恢复正常;在水稻中干涉OsMAX1a和OsMAX1e基因的表达能够增加水稻分蘖数。进一步的研究表明,外施GR24能够抑制OsMAX1a和OsMAX1e基因的表达,磷饥饿条件能够提高两个基因的表达水平从而促进独角金内酯的合成。另外,外施生长素和激动素能够显著影响OsMAX1a和OsMAX1e的表达。因此,OsMAX1a和OsMAX1e基因参与了独角金内酯的合成,并且在水稻生长发育中响应植物激素信号的诱导,是调控水稻分蘖的重要基因。

关键词: 独角金内酯, OsMAX1a, OsMAX1e, 分蘖

Abstract:

Strigolactones (SLs) are a class of phytohormone which play essential roles in regulating rice tillering, and ATMAX1 in Arabidopsis encodind P450 is an important gene involved in the synthesis of SLs.Our study found five MAX1 homologous genes through BLAST, namely OsMAX1a, OsMAX1b, OsMAX1c, OsMAX1d and OsMAX1e. Among them, OsMAX1a and OsMAX1e have stable and high expression in rice tissues. Our experiments verified that the overexpression of OsMAX1aand OsMAX1e in Arabidopsis rescued the max1 branched mutant phenotype, and the interference of OsMAX1a and OsMAX1e expression in rice increased rice tillering number. Furthermore, enogenous GR24 application inhibited the expression of OsMAX1a and OsMAX1e, and P-starvation enhanced the expression of OsMAX1a and OsMAX1e,thus promoted the biosynthesis of SLs. In addition, IAA and Kinetin(a cytokinin:KT) treatment apparently had remarkable effect on the expression of OsMAX1a and OsMAX1e. The results demonstrated that the expression of OsMAX1a and OsMAX1e responded to the SLs level and they were involved in the synthesis of SLs and regulated rice tillering.

Key words: strigolactones, OsMAX1a, OsMAX1e, tillering

中图分类号:

Q946
S511.01

王秀梅, 梁越洋, 李玲, 贡常委, 王海鹏, 黄晓西, 李双成, 邓其明, 朱军, 郑爱萍, 李平, 王世全. OsMAX1a, OsMAX1e通过参与独角金内酯的合成调控水稻分蘖[J]. 中国水稻科学, 2015, 29(3): 223-231.

Xiu-mei WANG, Yue-yang LIANG, Ling LI, Chang-wei GONG, Hai-peng WANG, Xiao-xi HUANG, Shuang-cheng LI, Qi-ming DENG, Jun ZHU, Ai-ping ZHENG, Ping LI, Shi-quan WANG. OsMAX1a and OsMAX1e, Involved in the Biosynthesis of Strigolactones,Regulate Rice Tillering[J]. Chinese Journal OF Rice Science, 2015, 29(3): 223-231.

图/表 9

图1 水稻OsMAX1候选基因在染色体上的位置及基因结构（B中黑色表示外显子, 细线表示内含子,空白区域为非编码区。）

Fig. 1. Chromosomal location and gene structure of rice OsMAX1 candidate genes.（In Fig. B, black boxes indicate exon and the connecting lines indicate intron.）

表1 水稻OsMAX1同源候选基因

Table 1 The OsMAX1 candidate genes in rice.

基因 Gene	登录号 ID	染色体 Chromosome	核苷酸大小 Nucleotide size / aa
OsMAX1a	LOC_Os01g50530	1	412
	LOC_Os01g50520	1
OsMAX1b	LOC_Os06g36920	6	549
OsMAX1c	LOC_Os01g50590	1	517
OsMAX1d	LOC_Os01g50580	1	385
OsMAX1e	LOC_Os02g12890	2	548

表2 本研究中用到的RNAi引物列表

Table 2 Sequence of RNAi markers used in the study.

引物 Marker	上游引物 Sence primer	下游引物 Anti-sence primer
RNAi-OsMAX1CON-F	5'-GCGTCGACGTTCTCAAGAGGCTTCGCTG-3'	5'-CGAGCTCGTTCTCAAGAGGCTTCGCTG-3'
RNAi-OsMAX1CON-R	5'-CCCAAGCTTCAAGGTAGGGGAATTTGGTCT-3'	5'-CGGAATTCCAAGGTAGGGGAATTTGGTCT-3'
RNAi-OsMAX1a-F	5'-GCGTCGAC TCCTGAAGGAAGCGATGAGA-3'	5'-CGAGCTC TCCTGAAGGAAGCGATGAGA-3'
RNAi-OsMAX1a-R	5'-CCCAAGCTTCACCTCTGGCTCTGGGAAAT-3'	5'-CGGAATTCCACCTCTGGCTCTGGGAAAT-3'
RNAi-OsMAX1e-F	5'-GCGTCGACGTCCAGCTCGCTGTCCACCA-3'	5'-TCCCCCGGGGTCCAGCTCGCTGTCCACCA-3'
RNAi-OsMAX1e-R	5'-CCCAAGCTTGAGACGAGGTGGAAGCGCATG-3'	5'-CGGAATTCGAGACGAGGTGGAAGCGCATG-3'

图2 OsMAX1 基因转录分析

Fig. 2. Transcriptional analysis of OsMAX1.

图3 拟南芥中OsMAX1a和OsMAX1e基因的过量表达（A,D-OsMAX1a基因过量表达转max1阳性植株T2代; B, E-OsMAX1e基因过表达转max1阳性植株T2代; C, F-野生型max1。）

Fig.3. Overexpression of OsMAX1a and OsMAX1e gene in Arabidopsis max1. （A and D, Over-expression of OsMAX1a in Arabidopsis max1; B and E, Over-expression of OsMAX1e in Arabidopsis max1; C and F, Arabidopsis max1.）

图4 水稻转基因植株阳性检测（+表示阳性对照(重组质粒);-表示阴性对照;1-12表示转基因植株。）

Fig. 4. PCR analysis of transgenic rice plants.（+, Positive control; -, Negative control; 1-12, Transgenic rice plants.）

图5 水稻 OsMAX1a、OsMAX1e干涉植株与日本晴分蘖数和株高比较（OsMAX1CON-RNAi为OsMAX1基因保守区域干涉。）

Fig. 5. Tiller number and height comparison of Nipponbare, OsMAX1a and OsMAX1e gene interference in rice.（OsMAX1CON-RNAi, The conserved region interference of OsMAX1 gene.）

图6 GR24和缺磷处理后OsMAX1a和OsMAX1e表达量变化

Fig. 6. Expression levels of OsMAX1a and OsMAX1e as affected by GR24 application and P-deficiency.

图 7 生长素(IAA)、激动素(KT)、N-1-萘氨甲酰苯甲酸(NPA)处理后OsMAX1a和OsMAX1e基因表达量变化

Fig. 7. Expression levels of OsMAX1a and OsMAX1e after IAA, KT, NPA treatment.

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