园艺学报 ›› 2021, Vol. 48 ›› Issue (1): 26-36.doi: 10.16420/j.issn.0513-353x.2020-0132
祁静静, 秦秀娟, 谢宇, 陈善春, 何永睿*(), 李强*(
)
收稿日期:
2020-06-05
修回日期:
2020-09-06
出版日期:
2021-01-25
发布日期:
2021-01-29
通讯作者:
何永睿,李强
E-mail:heyongrui@cric.cn;liqiang@cric.cn
基金资助:
QI Jingjing, QIN Xiujuan, XIE Yu, CHEN Shanchun, HE Yongrui*(), LI Qiang*(
)
Received:
2020-06-05
Revised:
2020-09-06
Online:
2021-01-25
Published:
2021-01-29
Contact:
HE Yongrui,LI Qiang
E-mail:heyongrui@cric.cn;liqiang@cric.cn
摘要:
为了探究过氧化氢酶基因CsKat01在柑橘抵御溃疡病过程中的作用,对感病品种‘晚锦橙’和抗病品种‘四季橘’中CsKat01进行了生物信息和表达分析,并探究了溃疡病菌接种处理后两品种中CAT酶活性和H2O2含量的关系。克隆CsKat01的编码序列并对该基因及其编码蛋白进行结构和功能分析,发现CsKat01编码框全长为1 482 bp,共编码493个氨基酸残基,蛋白序列中含有典型的CAT结构域;通过启动子序列克隆和分析发现‘晚锦橙’和‘四季橘’核心启动子区域均含有水杨酸、茉莉酸和脱落酸的响应元件,但数量不同;利用qRT-PCR分析水杨酸、茉莉酸、脱落酸和柑橘溃疡病菌(Xcc)对CsKat01的诱导表达特性,发现CsKat01的高表达可能使柑橘相对更感病;结合Xcc诱导后植物组织中CsKat01酶活性和H2O2含量,综合分析CsKat01、CAT酶活、H2O2含量和柑橘对溃疡病抗性之间的关系,推测CsKat01基因可能通过调控CAT的酶活性,改变H2O2含量进而影响柑橘对溃疡病的抗性。
中图分类号:
祁静静, 秦秀娟, 谢宇, 陈善春, 何永睿, 李强. 过氧化氢酶基因CsKat01与柑橘溃疡病相关性分析[J]. 园艺学报, 2021, 48(1): 26-36.
QI Jingjing, QIN Xiujuan, XIE Yu, CHEN Shanchun, HE Yongrui, LI Qiang. Correlation Analysis of Citrus Catalase Gene CsKat01 and Citrus Canker Disease[J]. Acta Horticulturae Sinica, 2021, 48(1): 26-36.
图2 多个物种过氧化氢酶蛋白序列比对 Cs:甜橙;Egl:桉树;Vv:葡萄;Nt:烟草;Pt:杨树;At:拟南芥。
Fig. 2 Multiple alignment of catalases of several organisms Cs:Citrus sinensis;Egl:Eucalyptus globulus;Vv:Vitis vinifera;Nt:Nicotiana tabacum;Pt:Populus trichocarpa;At:Arabidopsis thaliana.
图3 不同物种CAT的系统进化树 分支上的数字代表分支的可靠程度,长度代表进化距离。
Fig. 3 The phylogenetic tree of CAT from different species The number on the branch represents the reliability of the branch,and the length represents the evolutionary distance.
元件名称 Cis elements | 功能 Funtion | 序列 Sequence | 位置 Position | 方向 Strand | ||
---|---|---|---|---|---|---|
晚锦橙 Wanjincheng | 四季橘 Calamondin | 晚锦橙 Wanjincheng | 四季橘 Calamondin | |||
ABRE | 脱落酸响应 ABA-responsiveness | ACGTG | 733 | 733 | + | + |
1 536 | - | |||||
CCAAT-box | MYBHv1结合位点 MYBHv1 binding site | CAACGG | 1 307 | 1 320 | - | - |
CGTCA-motif | 茉莉酸响应 JA-responsiveness | CGTCA | 943 | 1 470 | + | - |
1 455 | - | |||||
TCA-element | 水杨酸响应元件 SA-responsiveness | CCATCTTTTT | 1 298 | 1 128 | - | + |
表1 Plant CARE预测CsKat01启动子区域顺式作用元件
Table 1 cis-Acting regulatory elements in promoter of CsKat01 predicted by Plant CARE
元件名称 Cis elements | 功能 Funtion | 序列 Sequence | 位置 Position | 方向 Strand | ||
---|---|---|---|---|---|---|
晚锦橙 Wanjincheng | 四季橘 Calamondin | 晚锦橙 Wanjincheng | 四季橘 Calamondin | |||
ABRE | 脱落酸响应 ABA-responsiveness | ACGTG | 733 | 733 | + | + |
1 536 | - | |||||
CCAAT-box | MYBHv1结合位点 MYBHv1 binding site | CAACGG | 1 307 | 1 320 | - | - |
CGTCA-motif | 茉莉酸响应 JA-responsiveness | CGTCA | 943 | 1 470 | + | - |
1 455 | - | |||||
TCA-element | 水杨酸响应元件 SA-responsiveness | CCATCTTTTT | 1 298 | 1 128 | - | + |
图4 水杨酸、茉莉酸和脱落酸诱导下CsKat01的相对表达 a、a’分别表示四季橘和晚锦橙的数据显著性差异,不同字母表示差异显著,P < 0.05。下同。
Fig. 4 The relative expression of CsKat01 induced by SA,JA and ABA The data significant difference of Calamondin and Wanjincheng is indicated by a and a’ respectively,and different letters indicate significant difference,P < 0.05. The same below.
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