Cloning and Expression Analysis of Two Key Genes of Jasmonic Acid Synthesis in Response to Endophytic Infection from Rehmannia glutinosa

doi:10.7525/j.issn.1673-5102.2021.02.017

Abstract

Abstract:

Allene oxide synthase（AOS） and 12-oxophytodienoate reductase（OPR） were the key genes of jasmonic acid biosynthesis in the plants. The genes of AOS and OPR responding to endophytic fungus infection were screened from the interaction transcriptome of Rehmannia glutinosa with endophytic fungus GG22. Specific primers were designed to the open reading frame of RgAOS and RgOPR. Bioinformatics analysis of the sequences and theirs encoded product were performed， and real-time fluorescent quantitative PCR was used to analyze the expression patterns of RgAOS and RgOPR in different tissues infection by endophytic fungus GG22. The open reading frame of RgAOS was 1 626 bp， encoding 541 amino acids with a molecular weight of 60.2 kD. The open reading frame of RgOPR was 1 197 bp， encoding 398 amino acids with a molecular weight of 44.07 kD. QPCR analysis showed that the expression of RgAOS was the highest in roots， the lowest in flowers， and the expression of RgOPR was higher in flowers and leaves， respectively. Endophytic fungus GG22 induced the expression of RgAOS and RgOPR in R.glutinosa. The RgAOS and RgOPR were successfully cloned from R.glutinosa， laying the foundation for further study on the biological activity of Jasmonic acid substances in R.glutinosa and claiming the molecular mechanism of endophytic fungi in the regulation of secondary metabolites of R.glutinosa.

Key words: allene oxide synthase, 12-oxophytodienoate reductase, bioinformatics analysis, expression analysis, Rehmannia glutinosa

CLC Number:

S567.9

Shu-Ping PENG, Cheng-Ming DONG, Yun-Hao ZHU. Cloning and Expression Analysis of Two Key Genes of Jasmonic Acid Synthesis in Response to Endophytic Infection from Rehmannia glutinosa[J]. Bulletin of Botanical Research, 2021, 41(2): 294-301.

References

1	朱畇昊，董诚明，李璐，等.地黄糖基转移酶基因克隆表达及表达分析［J］.中药材，2018，41（6）：1297-1301.
1	Zhu Y H，Dong C M，Li L，et al.Cloning and expression analysis of RgUGT gene in Rehmannia glutinosa［J］.Journal of Chinese Medicinal Materials，2018，41（6）：1297-1301.
2	刘彦飞，梁东，罗桓，等.地黄的化学成分研究［J］.中草药，2014，45（1）：16-22.
2	Liu Y F，Liang D，Luo H，et al.Chemical constituents from root tubers of Rehmannia glutinosa［J］.Chinese Traditional and Herbal Drugs，2014，45（1）：16-22.
3	张波泳，江振作，王跃飞，等.UPLC/ESI-Q-TOF MS法分析鲜地黄、生地黄、熟地黄的化学成分［J］.中成药，2016，38（5）：1104-1108.
3	Zhang B Y，Jiang Z Z，Wang Y F，et al.Analysis of chemical constituents in fresh，dried and prepared Rehmanniae Radix by UPLC/ESI-Q-TOF MS［J］.Chinese Traditional Patent Medicine，2016，38（5）：1104-1108.
4	Irmisch S，Jiang Y F，Chen F，et al.Terpene synthases and their contribution to herbivore-induced volatile emission in western balsam poplar（Populus trichocarpa）［J］.BMC Plant Biology，2014，14（1）：270.
5	Deavours B E，Dixon R A.Metabolic engineering of isoflavonoid biosynthesis in alfalfa［J］.Plant Physiology，2005，138（4）：2245-2259.
6	梁建萍，贾小云，刘亚令，等.干旱胁迫对蒙古黄芪生长及根部次生代谢物含量的影响［J］.生态学报，2016，36（14）：4415-4422.
6	Liang J P，Jia X Y，Liu Y L，et al.Effects of drought stress on seedling growth and accumulation of secondary metabolites in the roots of Astragalus membranaceus var.mongholicus［J］.Acta Ecologica Sinica，2016，36（14）：4415-4422.
7	Abouelsaad I，Renault S.Enhanced oxidative stress in the jasmonic acid-deficient tomato mutant，def-1，exposed to NaCl stress［J］.Journal of Plant Physiology，2018，226：136-144.
8	Jia H F，Zhang C，Pervaiz T，et al.Jasmonic acid involves in grape fruit ripening and resistant against Botrytis cinerea［J］.Functional & Integrative Genomics，2016，16（1）：79-94.
9	Fragoso V，Rothe E，Baldwin I T，et al.Root jasmonic acid synthesis and perception regulate folivore-induced shoot metabolites and increase Nicotiana attenuata resistance［J］.New Phytologist，2014，202（4）：1335-1345.
10	王云锋，李春琴，韩光煜，等.茉莉酸不同方式处理水稻对稻瘟病的防控效果及对水稻防御体系的影响［J］.南方农业学报，2019，50（3）：562-569.
10	Wang Y F，Li C Q，Han G Y，et al.Effect of different treatments of jasmonic acid on rice blast disease control and its effect on rice defense system［J］.Journal of Southern Agriculture，2019，50（3）：562-569.
11	Ren C G，Dai C C.Jasmonic acid is involved in the signaling pathway for fungal endophyte-induced volatile oil accumulation of Atractylodes lancea plantlets［J］.BMC Plant Biology，2012，12（1）：128.
12	冯莹莹，张钟秀，董先娟，等.白木香丙二烯氧化物合酶基因的表达分析［J］.药学学报，2017，52（12）：1962-1969.
12	Feng Y Y，Zhang Z X，Dong X J，et al.Expression analysis of allene oxide synthase gene from Aquilaria sinensis［J］.Acta Pharmaceutica Sinica，2017，52（12）：1962-1969.
13	Haga K，Lino M.Phytochrome-mediated transcriptional up-regulation of ALLENE OXIDE SYNTHASE in Rice Seedlings［J］.Plant and Cell Physiology，2004，45（2）：119-128.
14	吴洁，葛晓阳，李付广，等.陆地棉体细胞胚发育过程中AOS基因家族的表达谱初步分析及功能预测［J］.棉花学报，2016，28（6）：538-546.
14	Wu J，Ge X Y，Li F G，et al.Expression profile analysis and functional predictions of the AOS gene family in somatic embryos of upland cotton［J］.Cotton Science，2016，28（6）：538-546.
15	Mou Y F，Liu Y Y，Tian S J，et al.Genome-wide identification and characterization of the OPR gene family in wheat（Triticum aestivum L.）［J］.International Journal of Molecular Sciences，2019，20（8）：1914.
16	Stra?ner J，Fürholz A，Macheroux P，et al.A homolog of old yellow enzyme in tomato：spectral properties and substrate specificity of the recombinant protein［J］.Journal of Biological Chemistry，1999，274（49）：35067-35073.
17	Matsui H，Nakamura G，Ishiga Y，et al.Structure and expression of 12-oxophytodienoate reductase（subgroup I） genes in pea，and characterization of the oxidoreductase activities of their recombinant products［J］.Molecular Genetics and Genomics，2004，271（1）：1-10.
18	Zhang J L，Simmons C，Yalpani N，et al.Genomic analysis of the 12-oxo-phytodienoic acid reductase gene family of Zea mays［J］.Plant Molecular Biology，2005，59（2）：323-343.
19	Agrawal G K，Tamogami S，Han O，et al.Rice octadecanoid pathway［J］.Biochemical and Biophysical Research Communications，2004，317（1）：1-15.
20	吴娟娟，陈广通，李玉琴，等.金银花丙二烯氧化物合酶基因（LjAOS）的表达分析［J］.时珍国医国药，2012，23（4）：945-947.
20	Wu J J，Chen G T，Li Y Q，et al.Expression analysis of malondialdehyde synthase gene（LjAOS） in honeysuckle［J］.Lishizhen Medicine and Materia Medica Research，2012，23（4）：945-947.
21	王名雪，陆平，许菲，等.长春花丙二烯氧化物合酶基因的克隆与表达［J］.西北植物学报，2012，32（12）：2365-2373.
21	Wang M X，Lu P，Xu F，et al.Cloning and expression of allene oxide synthase gene from Catharanthus roseus［J］.Acta Botanica Boreali-Occidentalia Sinica，2012，32（12）：2365-2373.
22	蒋科技.药用植物茉莉酸合成途径关键酶基因的克隆与研究［D］.上海：复旦大学，2007.
22	Jiang K J.Cloning and studies of jasmonate biosynthetic pathway key genes from medicinal plants［D］.Shanghai：Fudan University，2007.
23	Lu X，Zhang F Y，Jiang W M，et al.Characterization of the first specific jasmonate biosynthetic pathway gene allene oxide synthase from Artemisia annua［J］.Molecular Biology Reports，2012，39（3）：2267-2274.
24	孙婷婷，王文举，刘峰，等.甘蔗茉莉酸合成关键酶基因ScOPR1的克隆、亚细胞定位及表达［J］.应用与环境生物学报，2018，24（6）：1365-1374.
24	Song T T，Wang W J，Liu F，et al.Molecular cloning，subcellular localization，and expression analysis of jasmonic acid synthesis gene ScOPR1 from sugarcane［J］.Chinese Journal of Applied and Environmental Biology，2018，24（6）：1365-1374.
25	游双红.葡萄VvXDH1、VvOPR1基因生物信息学分析及基因合成和功能鉴定［D］.南京：南京农业大学，2017.
25	You S H.Bioinformatics analysis，synthesis of VvXDH1，VvOPR1 in Vitis vinifera and functional identification［D］.Nanjing：Nanjing Agricultural University，2017.
26	付宇辰，刘晓慧，冷平生，等.金鱼草12-氧-植物二烯酸还原酶基因AmOPR的克隆及表达分析［J］.中国观赏园艺研究进展，2018，2018：504-510.
26	Fu Y C，Liu X H，Leng P S，et al.Gene cloning and expression analysis of AmOPR from Antirrhinum majus L.［J］.Advances in Ornamental Horticulture of China，2018，2018：504-510.
27	蔡昆争，董桃杏，徐涛.茉莉酸类物质（JAs）的生理特性及其在逆境胁迫中的抗性作用［J］.生态环境，2006，15（2）：397-404.
27	Cai K Z，Dong T X，Xu T.The physiological roles and resistance control in stress environment of jasmonates［J］.Ecology and Environment，2006，15（2）：397-404.
28	宋悦，崔晓山，陈娟娟，等.不同高温胁迫条件下的坛紫菜中植物激素分析［J］.水产学报，2017，41（10）：1578-1587.
28	Song Y，Cui X S，Chen J J，et al.The profiling of eleven phytohormones in Pyropia haitanensis under different high-temperature environments［J］.Journal of Fisheries of China，2017，41（10）：1578-1587.
29	Park J H，Halitschke R，Kim H B，et al.A knock‐out mutation in allene oxide synthase results in male sterility and defective wound signal transduction in Arabidopsis due to a block in jasmonic acid biosynthesis［J］.The Plant Journal，2002，31（1）：1-12.
30	Laudert D，Schsller F，Weiler E W.Transgenic Nicotiana tabacum and Arabidopsis thaliana plants overexpressing allene oxide synthase［J］.Planta，2000，211（1）：163-165.
31	连青龙，辛海波，李晓昕，等.唐菖蒲GhAOS基因的表达特性及其在拟南芥中的过表达分析［J］.中国农业科学，2012，45（14）：2923-2930.
31	Lian Q L，Xin H B，Li X X，et al.Analysis of GhAOS expression characteristics in Gladiolus hybridus and overexpression in Arabidopsis［J］.Scientia Agricultura Sinica，2012，45（14）：2923-2930.
32	Dolezal A L，Shu X M，Obrian G R，et al.Aspergillus flavus infection induces transcriptional and physical changes in developing maize kernels［J］.Frontiers in Microbiology，2014，5：384.
33	文锴，王远，胡蝶，等.不结球白菜BcOPR3基因的克隆与功能分析［J］.南京农业大学学报，2017，40（5）：804-811.
33	Wen K，Wang Y，Hu D，et al.Cloning and expression analysis of BcOPR3 gene in non-heading Chinese cabbage［J］.Journal of Nanjing Agricultural University，2017，40（5）：804-811.

[1]	Shanshan WANG, Rui WANG, Erqin FAN, Pengyue FU, Guanzheng QU, Nan WANG. Bioinformatics Analysis of CbuDELLAs Gene Family and Functional Analysis of CbuGRAS9 [J]. Bulletin of Botanical Research, 2024, 44(1): 139-151.
[2]	Chunyao WANG, Xiaojin LEI, Zhongyuan LIU. Expression Pattern Analysis of PdbHMGs Genes in Populus davidiana×P. bolleana under Abiotic Stress [J]. Bulletin of Botanical Research, 2023, 43(6): 932-942.
[3]	Shixian LIAO, Yuting WANG, Liben DONG, Yongmei GU, Fenglin JIA, Tingbo JIANG, Boru ZHOU. Function Analysis of the Transcription Factor PsnbZIP1 of Populus simonii×P. nigra in Response to Salt Stress [J]. Bulletin of Botanical Research, 2023, 43(2): 288-299.
[4]	Anying HUANG, Dean XIA, Yang ZHANG, Dongchen NA, Qing YAN, Zhigang WEI. Cloning and Drought Tolerance Expression Analysis of PtrWRKY51 Gene in Populus trichocarpa [J]. Bulletin of Botanical Research, 2022, 42(6): 1005-1013.
[5]	Huafeng CHEN, Longjun DAI, Mingyang LIU, Bingbing GUO, Hong YANG, Lifeng WANG. Stress Tolerance Functional Analysis of the High Expression Heat Shock Protein HbHSP90.4 Gene from the Latex of Hevea brasiliensis [J]. Bulletin of Botanical Research, 2022, 42(6): 1023-1032.
[6]	Denggao LI, Rui LIN, Qinghui MU, Na ZHOU, Yanru ZHANG, Wei BAI. Identification and Analysis of the Potato StCRKs Gene Family and Expression Patterns in Response to Stress Signals [J]. Bulletin of Botanical Research, 2022, 42(6): 1033-1043.
[7]	Mingyang LIU, Huaxing XIAO, Lifeng WANG, Xiaoxu LIANG, Yu ZHANG, Meng WANG. Cloning and Functional Analysis of Heat Shock Protein HbHSP90.8-1 from Hevea brasiliensis Müll. Arg. [J]. Bulletin of Botanical Research, 2022, 42(5): 811-820.
[8]	He CHENG, Shuanghui TIAN, Yang ZHANG, Cong LIU, De’an XIA, Zhigang WEI. Genome-wide Identification and Expression Analysis of nsLTP Gene Family in Populus trichocarpa [J]. Bulletin of Botanical Research, 2022, 42(3): 412-423.
[9]	Yuning Yang, Hao Dong, Shiwei Dong, Nairui Wang, Yue Song, Hanguo Zhang, Shujuan Li. Cloning and Expression Analysis of Transcription Factor LobHLH34 from Larix olgensis [J]. Bulletin of Botanical Research, 2022, 42(1): 112-120.
[10]	Jiaming Zhao, Erqin Fan, Yi Liu, Zhi Wang, Junhui Wang, Guanzheng Qu. Cloning and Bioinformatics Analysis of CbuATX1,CbuATX1-like and CbuATX2 Genes from Catalpa bungei [J]. Bulletin of Botanical Research, 2022, 42(1): 47-61.
[11]	Xin-Tong JI, Lei YU, Ya-Guang ZHAN. Cloning and Expression Analysis of BRASSINAZOLE RESISTANTANT1 (BZR1) Gene from Fraxinus mandshurica [J]. Bulletin of Botanical Research, 2021, 41(5): 744-752.
[12]	Xiao-Xiao WANG, Bi QIN, Yu-Shuang YANG, Qiu-Hai NIE, Ji-Chuan ZHANG, Shi-Zhong LIU. Cloning and Expression Analysis of E2 Ubiquitin-conjugating Enzyme Gene TkUBC2 in Taraxacum kok-saghyz Rodin [J]. Bulletin of Botanical Research, 2021, 41(1): 98-106.
[13]	Wen-Lin WANG, Hai-Sheng CHEN, Shu-Fang ZHENG, Song-Le FAN, Li-Feng WANG, Qiu-Jin TAN, Zhen-Shi QIN, Xi-Yun HUANG, Peng HE, Xiu-hua TANG, Peng XU. Cloning,Structure and Function Analysis of MiMYB2 Gene from Macadamia integrifolia [J]. Bulletin of Botanical Research, 2020, 40(6): 913-922.
[14]	WANG Wan-Qi, QI Wan-Zhu, ZHAO Qiu-Shuang, ZENG Dong, LIU Yi, FU Peng-Yue, QU Guan-Zheng, ZHAO Xi-Yang. Cloning and Expression Analysis of BpJMJ18 Gene Promoter in Betula platyphylla [J]. Bulletin of Botanical Research, 2020, 40(5): 751-759.
[15]	JIANG Cheng, ZHANG Xi, TIAN Qing, LI Li. Isolation of the BpbHLH112 Gene and Expression Analysis of Its Promoter in Betula platyphylla [J]. Bulletin of Botanical Research, 2020, 40(4): 583-592.