不同抗性烟草品系Rubisco及其活化酶对赤星病胁迫的响应

doi:10.13560/j.cnki.biotech.bull.1985.2023-0278

摘要/Abstract

摘要：

本研究以抗、感烟草品系JYH、CBH为试验材料，分析赤星病胁迫对光合作用和核酮糖-1,5-二磷酸核酮糖羧化酶/加氧酶（Rubisco）、核酮糖-1,5-二磷酸核酮糖羧化酶/加氧酶活化酶（RCA）活性及基因表达的影响。结果表明，胁迫9 d，JYH的光合作用及Rubisco、RCA活性、基因表达水平和蛋白含量受到明显抑制。赤星病胁迫显著降低CBH的光合作用及Rubisco、RCA活性和蛋白含量，Rubisco大、小亚基基因（rbcL、rbcS）、RCA基因（rca）的相对表达量也持续下降。JYH的Rubisco、RCA活性、基因表达水平和蛋白含量均显著高于CBH。相关性分析结果显示，P_n与Rubisco、RCA活性及基因表达量呈显著、极显著正相关。在赤星病胁迫下，JYH可维持较高的P_n与Rubisco、RCA活性，从而具有较强抗性。

关键词: 烟草赤星病, 光合作用, 1,5-二磷酸核酮糖羧化酶/加氧酶, 1,5-二磷酸核酮糖羧化酶/加氧酶活化酶, 基因表达, 相关性分析, 抗性差异, 品系

Abstract:

Using resistant and susceptible tobacco strains JYH and CBH as test materials, the effects of brown spot stress on the photosynthesis and activities of ribulose-1,5-bisphosphate carboxylase/oxygenase（Rubisco）and ribulose-1,5-diphosphate carboxylase/oxygenase activase（RCA）and their gene expressions were analyzed. The results showed that, there were obvious inhibition effects on a series of photosynthetic parameters and the activities, expression levels and protein contents of Rubisco and RCA in JYH at 9 d of brown spot stress. Meanwhile, tobacco brown spot stress notably reduced the photosynthesis and activities of Rubisco and RCA in CBH, in addition, the relative expression of Rubisco large and small subunits genes（rbcL and rbcS）and RCA genes（rca）and their protein contents all decreased under stress condition. In comparison with CBH, the activities, expression levels and protein contents of Rubisco and RCA in JYH were all significantly higher at the stress treatment of tobacco brown spot. The correlation analysis results also indicated that Pn had significant and extremely significant positive correlation with the activities, gene expression of Rubisco and RCA, respectively. These results suggest that the resistant strain JYH can maintain higher values of P_n, Rubisco and RCA activities under tobacco brown spot stress, which account for the resistance to brown spot stress.

Key words: tobacco brown spot, photosynthesis, Rubisco, RCA, gene expression, correlation analysis, resistant difference, strain

杨志晓, 侯骞, 刘国权, 卢志刚, 曹毅, 芶剑渝, 王轶, 林英超. 不同抗性烟草品系Rubisco及其活化酶对赤星病胁迫的响应[J]. 生物技术通报, 2023, 39(9): 202-212.

YANG Zhi-xiao, HOU Qian, LIU Guo-quan, LU Zhi-gang, CAO Yi, GOU Jian-yu, WANG Yi, LIN Ying-chao. Responses of Rubisco and Rubisco Activase in Different Resistant Tobacco Strains to Brown Spot Stress[J]. Biotechnology Bulletin, 2023, 39(9): 202-212.

图/表 6

表1 实时荧光定量PCR引物

Table 1 Primers for real-time quantitative RCR

基因 Gene	正向引物Forward primer（5'-3'）	反向引物Reverse primer（5'-3'）
rbcL	TTACAAAGGGCGATGCTACC	GAACCCAAATACGTTACCTACAATG
RbcS	GCTTCCTCTGTTCTTTCCTCTG	GCAATGGAAGTGATGTCAAGG
Rca	AGCAGCAGAAATCATCAG	ATTGTCCAGGGAAGGTAT
Actin	CCTGAGGTCCTTTTCCAACCA	GGATTCCGGCAGCTTCCATT

图1 赤星病胁迫对不同抗性烟草品系光合作用参数的影响 JYH-T和CBH-T：JYH、CBH接种烟草赤星病菌。JYH-CK和CBH-CK：JYH、CBH未接种烟草赤星病菌。不同字母表示差异显著（P<0.05）。下同

Fig. 1 Effects of brown spot stress on photosynthetic parameters in different resistant tobacco cultivars JYH-T and CBH-T: Inoculated with tobacco brown spot of JYH and CBH. JYH-CK and CBH-CK: Non-inoculated with tobacco brown spot of JYH and CBH. Different letters indicate significant differences（P<0.05）. The same below

图2 赤星病胁迫对不同抗性烟草品系Rubisco、RCA活性的影响

Fig. 2 Effects of brown spot stress on the activities of Rubisco and RCA in different resistant tobacco cultivars

图3 赤星病胁迫对不同抗性烟草品系rbcL、rbcS和rca相对表达量的影响

Fig. 3 Effects of brown spot stress on the relative expressions of rbcL, rbcS and rca in different resistant tobacco cultivars

图4 赤星病胁迫对不同抗性烟草品系rbcL、rbcS和rca含量的影响

Fig. 4 Effects of brown spot stress on the contents of rbcL, rbcS and rca in different resistant tobacco cultivars

表2 不同抗性烟草品系净光合速率与Rubisco、RCA基因表达特性的相关性分析

Table 2 Correlation analysis between net photosynthetic rate and gene expression characteristics of Rubisco and RCA in different resistant tobacco cultivars under brown spot stress

指标 Index	P_n	rbcL 相对表达量 rbcL relative expression	rbcS相对表达量 rbcS relative expression	rca相对表达量 rca relative expression	rbcL含量 Content of rbcL	rbcS含量 Content of rbcS	rca含量 Content of rca	Rubisco活化状态 Activation state of Rubisco	Rubisco 初始活性 Initial activity of Rubisco	Rubisco总活性 Total activity of Rubisco
rbcL相对表达量 rbcL relative expression	0.996**
rbcS相对表达量 rbcS relative expression	0.989**	0.427
rca相对表达量 rca relative expression	0.991**	0.193	0.704
rbcL含量 Content of rbcL	0.375	0.743	0.588	0.611
rbcS含量 Content of rbcS	0.442	0.347	-0.623	-0.194	0.927*
rca含量 Content of rca	0.296	0.204	0.037	0.974**	0.492	0.613
Rubisco活化状态 Activation state of Rubisco	0.926*	0.938*	0.198	0.427	0.128	0.327	0.979**
Rubisco初始活性 Initial activity of Rubisco	0.917*	0.962*	0.529	0.947*	-0.384	0.535	0.993**	0.584
Rubisco总活性 Total activity of Rubisco	0.943*	0.987**	-0.394	0.919*	0.456	0.136	0.982**	0.371	0.603
RCA活性 Activity of RCA	0.932*	0.449	0.438	0.996**	0.295	0.477	0.438	0.149	0.582	0.497

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