Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings

doi:10.3969/j.issn.1004-1524.2021.10.01

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (10): 1789-1796.DOI: 10.3969/j.issn.1004-1524.2021.10.01

Previous Articles Next Articles

Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings

LIU Han¹^,²(), DAI Yuanxing¹^,², LYU Mingfang², YUAN Zhengjie², LI Jing², YAN Chengqi², ZHANG Hengmu²^,^*()

1. College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China
2. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2021-04-15 Online:2021-10-25 Published:2021-11-02
Contact: ZHANG Hengmu

Abstract

Abstract:

To explore the effects of exogenous salicylic acid (SA) on plant growth and SA-related defense responses at the stage of rice seedlings, gradient concentrations of exogenous SA were used to spray rice seedlings of Nipponbare. Spectrophotometer was applied to measure the content of chlorophyll, a key indicator of rice seedling growth, and qRT-PCR was further used to analyze the expression levels of SA-biosynthesis gene PAL1, SA-receptor gene NPR1, and transcription factors WRKY45/WRKY76 and defense genes PR1a/PR1b at the downstream of SA signal pathway. It was found that different concentrations of exogenous SA exerted varying influences on expression patterns of tested genes, i.e. lower concentrations of exogenous SA promoted chlorophyll accumulation a certain extent and markedly affected on the expression of defense-related genes, while high concentrations of SA inhibited the accumulation of chlorophyll and induced abnormal growth at the stage of rice seedlings. Comprehensive comparison of quantitative results suggested that 2.0 mmol·L ^-1 of SA could be most effective in inducing the expression of defense-related genes by spraying rice seedlings. These results laid the foundation for further investigating roles of exogenous SA in promotion of plant growth and defense responses at the stage of rice seedlings.

Key words: rice, salicylic acid (SA), defense-related genes, chlorophyll

CLC Number:

S511.01

LIU Han, DAI Yuanxing, LYU Mingfang, YUAN Zhengjie, LI Jing, YAN Chengqi, ZHANG Hengmu. Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings[J]. Acta Agriculturae Zhejiangensis, 2021, 33(10): 1789-1796.

Figures/Tables 6

Table 1 Primers used in qPCR

目的基因 Target gene	上游引物(5'→3') Forward primer (5'→3')	下游引物(5'→3') Reverse primer (5'→3')	参考文献 Reference
OsNPR1	TTTCCGATGGAGGCAAGAG	GCTGTCATCCGAGCTAAGTGTT	[29]
OsPR1b	CAGCAACTGGAACAACCTTGG	TATGGACCGTGAAGGCGTGG	[25]
PAL1	CCGACCACCTGACTCACAA	ATCTCACGCTCGATGGACTT
OsPR1a	TATGCTATGCTACGTGTTTATGC	CACTAAGCAAATACGGCTGACA
OsWRKY45	TCCACGCGTGTGTACAGAAA	TGCTAGCATGTCTGCAGCTTA	[29]
OsWRKY76	CTGCCCGAATTCTAGCTTCCT	GCCCAAGGACCAACAGGTTAT
UBC	CCGTTTGTAGAGCCATAATTGCA	AGGTTGCCTGAGTCACAGTTAAGTG	[28]

Fig.1 Changes in chlorophyll content and phenotypes of rice seedlings caused by SA * represented significant differences at 0.05 level compared with 0 h. The same as below.

Fig.2 Relative expression level of OsPAL1 in SA-treated rice seedlings

Fig.3 Relative expression level of NPR1 in SA-treated rice seedlings

Fig.4 Relative expression levels of OsWRKY45 and OsWRKY76 in SA-treated rice seedlings

Fig.5 Relative expression levels of OsPR1a and OsPR1b in SA-treated rice seedlings

References 42

[1]	PENG Y J, VAN WERSCH R, ZHANG Y L. Convergent and divergent signaling in PAMP-triggered immunity and effector-triggered immunity[J]. Molecular Plant-Microbe Interactions, 2018, 31(4):403-409. DOI URL
[2]	RESJÖ S, ZAHID M A, BURRA D D, et al. Proteomics of PTI and two ETI immune reactions in potato leaves[J]. International Journal of Molecular Sciences, 2019, 20(19):4726. DOI URL
[3]	DING P T, DING Y L. Stories of salicylic acid: a plant defense hormone[J]. Trends in Plant Science, 2020, 25(6):549-565. DOI URL
[4]	JIANG G H, YIN D D, SHI Y, et al. OsNPR3.3-dependent salicylic acid signaling is involved in recessive gene xa5-mediated immunity to rice bacterial blight[J]. Scientific Reports, 2020, 10:6313. DOI URL
[5]	DUAN L, LIU H B, LI X H, et al. Multiple phytohormones and phytoalexins are involved in disease resistance to Magnaporthe oryzae invaded from roots in rice[J]. Physiologia Plantarum, 2014, 152(3):486-500. DOI URL
[6]	RIVAS-SAN VICENTE M, PLASENCIA J. Salicylic acid beyond defence: its role in plant growth and development[J]. Journal of Experimental Botany, 2011, 62(10):3321-3338. DOI URL
[7]	BETSUYAKU S, KATOU S, TAKEBAYASHI Y, et al. Salicylic acid and jasmonic acid pathways are activated in spatially different domains around the infection site during effector-triggered immunity in Arabidopsis thaliana[J]. Plant and Cell Physiology, 2018, 59(1):8-16. DOI URL
[8]	SHEN C J, YANG Y J, LIU K D, et al. Involvement of endogenous salicylic acid in iron-deficiency responses in Arabidopsis[J]. Journal of Experimental Botany, 2016, 67(14):4179-4193. DOI URL
[9]	DE VLEESSCHAUWER D, GHEYSEN G, HÖFTE M. Hormone defense networking in rice: tales from a different world[J]. Trends in Plant Science, 2013, 18(10):555-565. DOI URL
[10]	SILVERMAN P, SESKAR M, KANTER D, et al. Salicylic acid in rice (biosynjournal, conjugation, and possible role)[J]. Plant Physiology, 1995, 108(2):633-639. DOI URL
[11]	YALPANI N, SILVERMAN P, WILSON T M, et al. Salicylic acid is a systemic signal and an inducer of pathogenesis-related proteins in virus-infected tobacco[J]. The Plant Cell, 1991, 3(8):809-818.
[12]	LEMARIÉ S, ROBERT-SEILANIANTZ A, LARIAGON C, et al. Both the jasmonic acid and the salicylic acid pathways contribute to resistance to the biotrophic clubroot agent Plasmodiophora brassicae in Arabidopsis[J]. Plant and Cell Physiology, 2015, 56(11):2158-2168.
[13]	ENYEDI A J, YALPANI N, SILVERMAN P, et al. Localization, conjugation, and function of salicylic acid in tobacco during the hypersensitive reaction to tobacco mosaic virus[J]. Proceedings of the National Academy of Sciences of USA, 1992, 89(6):2480-2484. DOI URL
[14]	LOVE A J, GERI C, LAIRD J, et al. Cauliflower mosaic virus protein P6 inhibits signaling responses to salicylic acid and regulates innate immunity[J]. PLoS One, 2012, 7(10):e47535. DOI URL
[15]	ZHANG W N, CHEN W L. Role of salicylic acid in alleviating photochemical damage and autophagic cell death induction of cadmium stress in Arabidopsis thaliana[J]. Photochemical & Photobiological Sciences, 2011, 10(6):947-955.
[16]	LOVELOCK D A, ŠOLA I, MARSCHOLLEK S, et al. Analysis of salicylic acid-dependent pathways in Arabidopsis thaliana following infection with Plasmodiophora brassicae and the influence of salicylic acid on disease[J]. Molecular Plant Pathology, 2016, 17(8):1237-1251. DOI URL
[17]	WHITE R F. Acetylsalicylic acid (aspirin) induces resistance to tobacco mosaic virus in tobacco[J]. Virology, 1979, 99(2):410-412. DOI URL
[18]	CHEN Z, CHEN T, SATHE A, et al. Identification of a novel semi-dominant spotted-leaf mutant with enhanced resistance to Xanthomonas oryzae pv. oryzae in rice[J]. International Journal of Molecular Sciences, 2018, 19(12):3766. DOI URL
[19]	STELLA DE FREITAS T F, STOUT M J, SANT'ANA J. Effects of exogenous methyl jasmonate and salicylic acid on rice resistance to Oebalus pugnax[J]. Pest Management Science, 2019, 75(3):744-752. DOI URL
[20]	FENG B H, ZHANG C X, CHEN T T, et al. Salicylic acid reverses pollen abortion of rice caused by heat stress[J]. BMC Plant Biology, 2018, 18(1):245. DOI URL
[21]	LOPES F B, SANT’ANA J. Responses of Spodoptera frugiperda and Trichogramma pretiosum to rice plants exposed to herbivory and phytohormones[J]. Neotropical Entomology, 2019, 48(3):381-390. DOI URL
[22]	MOU S L, SHI L P, LIN W, et al. Over-expression of rice CBS domain containing protein, OsCBSX3, confers rice resistance to Magnaporthe oryzae inoculation[J]. International Journal of Molecular Sciences, 2015, 16(7):15903-15917. DOI URL
[23]	PENG X X, WANG H H, JANG J C, et al. OsWRKY80-OsWRKY4 module as a positive regulatory circuit in rice resistance against Rhizoctonia solani[J]. Rice, 2016, 9(1):1-14. DOI URL
[24]	LIU H, DONG S Y, SUN D Y, et al. CONSTANS-like 9 (OsCOL9) interacts with receptor for activated C-kinase 1(OsRACK1) to regulate blast resistance through salicylic acid and ethylene signaling pathways[J]. PLoS One, 2016, 11(11):e0166249. DOI URL
[25]	XIE X Z, XUE Y J, ZHOU J J, et al. Phytochromes regulate SA and JA signaling pathways in rice and are required for developmentally controlled resistance to Magnaporthe grisea[J]. Molecular Plant, 2011, 4(4):688-696. DOI URL
[26]	李燕, 龙湍, 吴昌银. 组培水稻种子发芽[EB/OL].(2018-03-16)[2021-04-15]. Bio-101, 2018: e1010183. DOI: 10.21769/BioProtoc.1010183.
	LI Y, LONG T, WU C Y. Germination of rice seeds with tissue culture method [EB/OL].(2018-03-16)[2021-04-15]. Bio-101, 2018: e1010183. DOI: 10.21769/BioProtoc.1010183.
[27]	徐春梅, 陈丽萍, 王丹英, 等. 低氧胁迫对水稻幼苗根系功能和氮代谢相关酶活性的影响[J]. 中国农业科学, 2016, 49(8):1625-1634.
	XU C M, CHEN L P, WANG D Y, et al. Effects of low oxygen stress on the root function and enzyme activities related to nitrogen metabolism in roots of rice seedlings[J]. Scientia Agricultura Sinica, 2016, 49(8):1625-1634.(in Chinese with English abstract)
[28]	项聪英, 蔡年俊, 李静, 等. 一个水稻小热休克蛋白基因的克隆和鉴定[J]. 中国水稻科学, 2016, 30(6):587-592.
	XIANG C Y, CAI N J, LI J, et al. Cloning and characterization of a small heat shock protein (SHSP) gene in rice plant[J]. Chinese Journal of Rice Science, 2016, 30(6):587-592.(in Chinese with English abstract)
[29]	SHIMONO M, SUGANO S, NAKAYAMA A, et al. Rice WRKY45 plays a crucial role in benzothiadiazole-inducible blast resistance[J]. The Plant Cell, 2007, 19(6):2064-2076. DOI URL
[30]	周勇, 范晓磊, 林拥军, 等. 水稻叶绿素含量的测定[EB/OL].(2018-06-12)[2021-04-15]. Bio-101, 2018: e1010147.DOI: 10.21769/BioProtoc.1010147.
	ZHOU Y, FAN X L, LIN Y J, et al. Determination of chlorophyll content in rice[EB/OL].(2018-06-12)[2021-04-15]. Bio-101, 2018: e1010147. DOI: 10.21769/BioProtoc.1010147.
[31]	DEMPSEY D A, VLOT A C, WILDERMUTH M C, et al. Salicylic acid biosynjournal and metabolism[J]. The Arabidopsis Book, 2011, 9:e0156. DOI URL
[32]	PAJEROWSKA-MUKHTAR K M, EMERINE D K, MUKHTAR M S. Tell me more: roles of NPRs in plant immunity[J]. Trends in Plant Science, 2013, 18(7):402-411. DOI URL
[33]	DING Y L, SUN T J, AO K, et al. Opposite roles of salicylic acid receptors NPR1 and NPR3/NPR4 in transcriptional regulation of plant immunity[J]. Cell, 2018, 173(6):1454-1467. DOI URL
[34]	ZHANG Y L, LI X. Salicylic acid: biosynjournal, perception, and contributions to plant immunity[J]. Current Opinion in Plant Biology, 2019, 50:29-36. DOI URL
[35]	RYU H S, HAN M, LEE S K, et al. A comprehensive expression analysis of the WRKY gene superfamily in rice plants during defense response[J]. Plant Cell Reports, 2006, 25(8):836-847. DOI URL
[36]	LIANG X X, CHEN X J, LI C, et al. Metabolic and transcriptional alternations for defense by interfering OsWRKY62 and OsWRKY76 transcriptions in rice[J]. Scientific Reports, 2017, 7:2474. DOI URL
[37]	HUANGFU J Y, LI J C, LI R, et al. The transcription factor OsWRKY45 negatively modulates the resistance of rice to the brown planthopper Nilaparvata lugens[J]. International Journal of Molecular Sciences, 2016, 17(6):697. DOI URL
[38]	PENG Y, BARTLEY L E, CANLAS P, et al. OsWRKY IIa transcription factors modulate rice innate immunity[J]. Rice, 2010, 3(1):36-42. DOI URL
[39]	LIU X Y, ROCKETT K S, KØRNER C J, et al. Salicylic acid signalling: new insights and prospects at a quarter-century milestone[J]. Essays in Biochemistry, 2015, 58:101-113. DOI URL
[40]	KU Y S, SINTAHA M, CHEUNG M Y, et al. Plant hormone signaling crosstalks between biotic and abiotic stress responses[J]. International Journal of Molecular Sciences, 2018, 19(10):3206. DOI URL
[41]	MUR L A J, KENTON P, ATZORN R, et al. The outcomes of concentration-specific interactions between salicylate and jasmonate signaling include synergy, antagonism, and oxidative stress leading to cell death[J]. Plant Physiology, 2006, 140(1):249-262. DOI URL
[42]	DEWEZ D, DAUTREMEPUITS C, JEANDET P, et al. Effects of methanol on photosynthetic processes and growth of Lemna gibba[J]. Photochemistry and Photobiology, 2003, 78(4):420-424. DOI URL

Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 42

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	PEI Huimin, WU Mingming, ZHAI Rongrong, YE Jing, JIN Yue, ZHU Yi, HOU Jianjun, ZHU Guofu, YE Shenghai. Research progress on gene function and breeding of low-cadmium rice cultivars [J]. Acta Agriculturae Zhejiangensis, 2025, 37(9): 2012-2020.
[2]	TAN Shiyi, YU Guohong, XUE Xianglei, ZHAO Yinglei, XU Baoyu, ZHANG Chenghao. Design and experiment of tray handling device for industrialized rice seedling raising [J]. Acta Agriculturae Zhejiangensis, 2025, 37(7): 1545-1555.
[3]	ZHANG Zhi, HE Haohao, YU Miao, XU Jianfeng. Effects of chemical fertilizer reduction combined with soil conditioner on soil acidity, soil nutrients and rice yield [J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1301-1308.
[4]	LIN Xiaobing, LI Jiang, CHENG Yanhong, WANG Binqiang, HE Shaolang, HUANG Shangshu, HUANG Qianru. Effects of organic materials on soil microbial biomass, mineral nitrogen content and rice yield [J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1309-1318.
[5]	SU Yang, SHANG Xiaolan, QIAN Zhongming, WU Lingen, HUANG Jiaqi, ZHUANG Haifeng, ZHAO Yufei, DANG Hongyang, XU Lijun. Effects of synergistic enhancement of straw returning to the field with decomposition agent and biochar on soil quality and rice growth [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 1139-1148.
[6]	ZHU Xiao, ZHU Ying, LI Hongjun, CHEN Shanfeng. Preparation and quality of oat chickpea compound rice by squeezing [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 1149-1158.
[7]	LIU Qihua, SUN Zhaowen, ZHENG Chongke. Effects of nitrogen management on absorption and allocation of microelements in above-ground parts of dry direct-sowing rice [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 987-997.
[8]	YING Yongfei, HAN Dongxuan, MENG Fang, YU Lin, SHEN Jialuan, WANG Kaiying. Effects of biogas slurry substituting chemical fertilizer on rice yield and quality and soil characteristics [J]. Acta Agriculturae Zhejiangensis, 2025, 37(4): 880-891.
[9]	SONG Xinlu, FAN Shuhong, WU Guangqi, ZHAN Mengqi, HOU Qian, LI Mingyue, XU Yan. Effects of combined copper-phenanthrene pollution on physiological characteristics and pollutant accumulation of rice roots at tillering stage [J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 521-529.
[10]	LEI Zhiwei, LI Xinxin, XU Heng, ZHANG Heng, ZHU Ying, ZHANG Hua. Identification of QTLs for stem borer resistance using chromosome segment substitution lines in rice [J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 530-537.
[11]	XIE Changyan, JIN Yumeng, ZHANG Miao, DONG Qingjun, LI Qing, JI Li, ZHONG Ping, CHEN Chuan, ZHANG Ankang. Application effect of nutrient soil made from river sludge for machine-transplanted rice seedling [J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 538-547.
[12]	LI Guiping, XU Xiaomei, LU Lizhi. Rice-duck farming and its environmental effects [J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 643-653.
[13]	WU Jiaqi, ZHU Xueming, BAO Jiandong, WANG Caoyi, ZHOU Xiaoyu, LI Lin, LIN Fucheng. Research progress on biological control of rice blast [J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 736-744.
[14]	WAN Shaoyuan, LIU Xianbo, CAI Shuo, SHI Hong, CHENG Jie. Effect of irrigation and planting methods on the yield and quality of double-cropping rice [J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 257-268.
[15]	LAN Xuecheng, ZHAO Fengliang, ZHANG Guangxu, LI Yang, GUO Xiaohong. Effects of nano zinc oxide and nano silicon dioxide on rice seed germination [J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 269-277.