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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (6): 1518-1531.doi: 10.3724/SP.J.1006.2023.24153

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Effects of BnMAPK2 on drought tolerance in Brassica napus

YUAN Da-Shuang1,2,3(), ZHANG Xiao-Li1,2, ZHU Dong-Ming1,2, YANG You-Hong1,2, YAO Meng-Nan1,2, LIANG Ying1,2,*()   

  1. 1College of Agronomy and Biotechnology, Southwest University/Chongqing Engineering Research Center for Rapeseed, Chongqing 400715, China
    2Academy of Agricultural Sciences, Southwest University, Chongqing 400715, China
    3Natural Resources Bureau of Liuzhi Special Economic Zone, Liupanshui City, Guizhou Province, Liupanshui 553400, Guizhou, China
  • Received:2022-07-01 Accepted:2022-10-10 Online:2023-06-12 Published:2023-04-07
  • Contact: *E-mail: yliang@swu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(31872876)

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Abstract:

The mitogen-activated protein kinase (MAPK) cascade is involved in various biotic and abiotic stress responses in plants. BnMAPK2 belongs to the most downstream C-family genes in the MAPK cascade pathway. In this study, BnMAPK2 overexpression (OE-MAPK2) and RNA interference expression (RNAi-MAPK2) transgenic Brassica napus were successfully obtained. Under drought conditions, the drought tolerance of OE-MAPK2 plants was increased, and the drought tolerance of RNAi-MAPK2 plants was decreased. The related physiological indicators showed that under drought stress BnMAPK2 could slow down the degree of leaf dehydration, promote the accumulation of proline in plants, reduce the content of malondialdehyde, and increase the activity of POD at the later stage of drought. We compared the differences in the relative expression levels between transgenic and wild-type plants in the drought-related genes (P5CSB, SCE1), BnMAPK2-interacting drought-related genes (STRS2, CRL1), and STRS2-dependent ABA signaling pathway-related genes (RD22, MYC, SnRK2). The results showed that BnMAPK2 positively regulated the relative expression levels of P5CSB, SCE1, CRL1, RD22, MYC, and SnRK2, negatively regulated the relative expression levels of STRS2, and the relative expression trends of genes related to STRS2-dependent ABA signaling pathway in OE-MAPK2 plants and strs2 mutants were consistent. Therefore, it was speculated that BnMAPK2 can increase plant drought tolerance by regulating the in vivo permeability, leaf water content, cell membrane and protein structure stability, scavenging free radicals, and reducing membrane lipid peroxidation, which can also negatively regulate STRS2 by interacting with STRS2. The relative expression of the genes mediated the STRS2-dependent ABA signaling pathway and increased the drought tolerance of plants. This study lays a foundation for further elucidating the anti-stress mechanism of BnMAPK2 gene.

Key words: Brassica napus, BnMAPK2, drought tolerance, qRT-PCR

Table 1

Primers used in this study"

引物名称
Primer name		 上游引物
Forward sequence (5′-3′)		 下游引物
Reverse sequence (5′-3′)
OV-BnMAPK2 CACCATGTTATTGCATAACTTGTCTGAAGG GAGCTCAGAGTTAACAGTTTCTGGA
impk2-A ctGACGTCTATTCCCGGGGACAGAATGTC cgCCATGGTCTGAAGCAGATCAATGGCTAG
impk2-B ctGGATCCTATTCCCGGGGACAGAATGTC cgTCTAGATCTGAAGCAGATCAATGGCTAG
F35s3ND GGAAGTTCATTTCATTTGGAGAG GCTGCATAATTCTCGGGGCAGCA
Bar CGACATCCGCCGTGCCACCGA CAAATCTCGGTGACGGGCAGG
26S CACAATGATAGGAAGAGCCGAC CAAGGGAACGGGCTTGGCAGAATC
ACT7 TGGGTTTGCTGGTGACGAT TGCCTAGGACGACCAACAATACT
MAPK2-q GGGGACAGAATGTCTTAACCAGA GGGAGAGACTCTATGTATCTTTTG
P5CSB CAGAAGCCACAGACTGAACTTG AAACTGCTATCAGTCACCAGCA
PLC CTGATCGATGTTCAGAAGCAAG TCGAGGTGGAGACCGTTACTAT
SCE1 AGGCTTTTTCCACCCTAATGTCTATCCA ACCCTTTTCTTGTACTCAACTGCATCC
STRS2 GAAGCAAGATCATGAGTTCGTC CTGGTTCTGTAACTCTGAGGTT
SnRK ATATCGAGCGAGGTGAGAAGA AGCTGCGTATTCCATAACAATA
BnMYC2 CGACGATAACGCCTCTATGA CCTTCGTTTGTCCCTTCAAT
BnRD22 CGCAGCGGCTGGAGTAAAGA ACCGCGTAAACGCTCGTCAT

Fig. 1

Identification of the relative expression levels of BnMAPK2 genes in transgenic plants and wild-type plants Different lowercase letters are significantly different at P < 0.05 within the same treatment."

Fig. 2

Relative expression levels of BnMAPK2 genes under drought conditions Data are presented as means ± SDs (n = 3)."

Fig. 3

Comparison of leaf water content and phenotype between transgenic and wild-type plants Data are presented as means ± SDs (n = 3). *, **, and *** indicate significant differences at the 0.05, 0.01, and 0.001 probability levels, respectively. Significance refers to the difference between the transgenic plant and the wild type."

Fig. 4

Comparison of proline and malondialdehyde between transgenic and wild-type plants under natural drought conditions Data are presented as means ± SDs (n = 3). *, **, and *** indicate significant differences at the 0.05, 0.01, and 0.001 probability levels, respectively. Significance refers to the difference between the transgenic plant and the wild type."

Fig. 5

Comparison of antioxidant enzyme contents in transgenic plants and wild-type plants under natural drought conditions Data are presented as means ± SDs (n = 3). *, **, and *** indicate significant differences at the 0.05, 0.01, and 0.001 probability levels, respectively. Significance refers to the difference between the transgenic plant and the wild type."

Fig. 6

Relative expression levels of drought-related genes in transgenic and wild-type plants under drought conditions Data are presented as means ± SDs (n = 3). *, **, and *** indicate significant differences at the 0.05, 0.01, and 0.001 probability levels, respectively. Significance refers to the difference between the transgenic plant and the wild type."

Fig. 7

Relative expression levels of BnMAPK2 interacting genes under drought conditions Data are presented as means ± SDs (n = 3). *, **, and *** indicate significant differences at the 0.05, 0.01, and 0.001 probability levels, respectively. Significance refers to the difference between the transgenic plant and the wild type."

Fig. 8

Gene expression patterns related to ABA pathway Data are presented as mean ± SD (n = 3). *, **, and *** indicate significant differences at the 0.05, 0.01, and 0.001 probability levels, respectively. Significance refers to the difference between the transgenic plant and the wild type."

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[1] Li Shaoqing, Li Yangsheng, Wu Fushun, Liao Jianglin, Li Damo. Optimum Fertilization and Its Corresponding Mechanism under Complete Submergence at Booting Stage in Rice[J]. Acta Agronomica Sinica, 2002, 28(01): 115 -120 .
[2] Wang Lanzhen;Mi Guohua;Chen Fanjun;Zhang Fusuo. Response to Phosphorus Deficiency of Two Winter Wheat Cultivars with Different Yield Components[J]. Acta Agron Sin, 2003, 29(06): 867 -870 .
[3] YANG Jian-Chang;ZHANG Jian-Hua;WANG Zhi-Qin;ZH0U Qing-Sen. Changes in Contents of Polyamines in the Flag Leaf and Their Relationship with Drought-resistance of Rice Cultivars under Water Deficiency Stress[J]. Acta Agron Sin, 2004, 30(11): 1069 -1075 .
[4] Yan Mei;Yang Guangsheng;Fu Tingdong;Yan Hongyan. Studies on the Ecotypical Male Sterile-fertile Line of Brassica napus L.Ⅲ. Sensitivity to Temperature of 8-8112AB and Its Inheritance[J]. Acta Agron Sin, 2003, 29(03): 330 -335 .
[5] Wang Yongsheng;Wang Jing;Duan Jingya;Wang Jinfa;Liu Liangshi. Isolation and Genetic Research of a Dwarf Tiilering Mutant Rice[J]. Acta Agron Sin, 2002, 28(02): 235 -239 .
[6] WANG Li-Yan;ZHAO Ke-Fu. Some Physiological Response of Zea mays under Salt-stress[J]. Acta Agron Sin, 2005, 31(02): 264 -268 .
[7] TIAN Meng-Liang;HUNAG Yu-Bi;TAN Gong-Xie;LIU Yong-Jian;RONG Ting-Zhao. Sequence Polymorphism of waxy Genes in Landraces of Waxy Maize from Southwest China[J]. Acta Agron Sin, 2008, 34(05): 729 -736 .
[8] HU Xi-Yuan;LI Jian-Ping;SONG Xi-Fang. Efficiency of Spatial Statistical Analysis in Superior Genotype Selection of Plant Breeding[J]. Acta Agron Sin, 2008, 34(03): 412 -417 .
[9] WANG Yan;QIU Li-Ming;XIE Wen-Juan;HUANG Wei;YE Feng;ZHANG Fu-Chun;MA Ji. Cold Tolerance of Transgenic Tobacco Carrying Gene Encoding Insect Antifreeze Protein[J]. Acta Agron Sin, 2008, 34(03): 397 -402 .
[10] ZHENG Xi;WU Jian-Guo;LOU Xiang-Yang;XU Hai-Ming;SHI Chun-Hai. Mapping and Analysis of QTLs on Maternal and Endosperm Genomes for Histidine and Arginine in Rice (Oryza sativa L.) across Environments[J]. Acta Agron Sin, 2008, 34(03): 369 -375 .
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