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| Identification of NAC Family Genes in Cajanus cajan and Analysis of Their Response to Fungal Infection |
| ZHANG Xiu-Qi1*, WU Rui1*, DONG Bi-Ying1, DU Ting-Ting1, SONG Zhi-Hua1, LI Na1, CAO Hong-Yan1, YANG Qing1,2, MENG Dong1,2** |
1 The Key Laboratory for Silviculture and Conservation of Ministry of Education, College of Forestry, Beijing Forestry University, Beijing 100083, China;
2 Institute of Tree Development and Gene Editing, Beijing Forestry University, Beijing 100083, China |
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Abstract The NAC (NAM, ATAF1/2 and CUC2) transcription factor family is a collective name for genes with conserved NAC structural domains that play key regulatory roles in plant growth, development, and stress response and other life activities. In this study, 90 CcNAC candidate genes were initially identified from the Cajanus cajan genome by sequence alignment using the NAC family genes of Arabidopsis thaliana. Phylogenetic analysis showed that the 90 CcNAC genes could be divided into 13 subfamilies, and a total of 24 CcNAC family genes were screened as potential biotic stress response genes in subfamilies Group1~4 based on the study of Arabidopsis gene functions. Further physicochemical characterization and cis-acting element analysis of the 24 CcNAC family genes showed that the promoter region contains several phytohormone response elements, which might be closely related to hormone signaling and have potential responses to biotic and abiotic stresses function. Transcriptome analysis showed that 13 NAC genes were significantly up- regulated and 5 NAC genes were significantly down-regulated at 12 h of treatment under the effect of MeJA, among which CcNAC55 was significantly up-regulated at 3, 6 and 12 h of treatment, and the highest expression levels were found at 6 and 12 h. Further, using semi-quantitative assays, CcNAC55 was found that it was sustaining significantly up-regulated under fungal Cc1-1 stress, while there was no significant difference under high temperature stress, suggested that CcNAC55 might play an important role in the response of Cajanus cajan to biotic stress. To verify the function of CcNAC55 in response to biotic stress, in this study, the lines of Cajanus cajan overexpressed CcNAC55 were established by vacuum osmotic transformation and inoculated the leaves with the pathogenic fungus Cc1-1. The results showed that CcNAC55 overexpression could significantly reduce the infestation rate and proportion of infected area of Cajanus cajan leaves. This study provides a reference for the study of NAC gene function in Cajanus cajan and will help to identify and select candidate genes related to biotic stress tolerance.
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Received: 11 June 2022
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Corresponding Authors:
**mengdongjlf@163.com
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| About author:: * These authors contributed equally to this work |
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[1] 杜婷婷, 宋治华, 董碧莹, 等. 2021. 木豆类黄酮代谢通路关键基因家族的鉴定与表达分析[J]. 农业生物技术学报, 29(12): 2289-2303.
(DU T T, Song Z H, Dong B Y, et al. 2021. ldentification and expression analysis of key gene families in flavonoid metabolism pathway in pigeon pea (Cajanus cajan)[J]. Journal of Agricultural Biotechnology, 29(12): 2289-2303.)
[2] 李娜, 宋治华, 范雨欣, 等. 2021. 木豆 JAZ 基因家族鉴定及其响应致病真菌 Cc1-1 侵染的表达分析[J]. 农业生物技术学报, 29(08): 1495-1505.
(Li N, Song Z H, Fan Y X, et al. 2021. Identification of JAZ gene family in Caja-nus cajan and expression analysis in response to pathogenic fungus Cc1-1[J]. Journal of Agricultural Biotechnology, 29(08): 1495-1505.)
[3] 彭宇婧, 吕若岚, 李育军, 等. 2021. 木豆加工与综合利用现状 、展望及产业发展建议[J]. 长江蔬菜, 20: 37-40.
(Peng Y J, Lv R L, Li Y J, et al. Current status, prospect and industrial development suggestions on processing and comprehensive utilization of pigeonpea[J]. Journal of Changjiang Vegetables, (20): 37-40.)
[4] 周文楠, 郭志鹏, 牛军鹏, 等. 2019. 外源茉莉酸甲酯对紫花苜蓿尖孢镰刀菌根腐病抗病性的作用[J]. 植物病理学报, 49(03): 379-390.
(Zhou W N, Guo Z P, Niu J P, et al. Effect of methyl jasmonate on resistance of alfalfa root rot caused by Fusarium oxysporum[J]. Acta Phytopathologica Sinica, 49(03): 379-390.)
[5] Bohra A, Saxena K B, Varshney R K, et al. 2020. Genomics- ass-isted breeding for pigeonpea improvement[J]. Theoretical and Applied Genetics, 133(5): 1721-1737.
[6] Bu Q, Jiang H, Li C B, et al. 2008. Role of the Arabidopsis thaliana NAC transcription factors ANAC019 and ANAC055 in regulating jasmonic acid-signaled defense responses[J]. Cell Research, 18(7): 756-767.
[7] Delessert C, Kazan K, Wilson I W, et al. 2005. The transcription factor ATAF2 represses the expression of pathogenesis-related genes in Arabidopsis[J]. The Plant Journal, 43(5): 745-757
[8] Diao W, Snyder J C, Wang S, et al. 2018. Genome-wide analyses of the NAC transcription factor gene family in pepper (Capsicum annuum L.): Chromosome location, phylogeny, structure, expression patterns, cis-elements in the promoter, and interaction network[J]. International Journal of Molecular Sciences, 19(4): 1048-1461.
[9] Du T T, Fan Y X, Cao H Y, et al. 2021. Transcriptome analysis revealed key genes involved in flavonoid metabolism in response to jasmonic acid in pigeon pea (Caja- nus cajan (L.) Millsp.)[J]. Plant Physiology and Biochemistry, 168: 410-422.
[10] Duval M, Hsieh T F, Kim S Y, et al. 2002. Molecular characterization of AtNAM: A member of the Arabidopsis NAC domain superfamily[J]. Plant Molecular Biology, 50(2): 237-248.
[11] Fang Y, You J, Xie K, et al. 2008. Systematic sequence analysis and identification of tissue-specific or stress-responsive genes of NAC transcription factor family in rice[J]. Molecular Genetics and Genomics, 280(6): 547-63.
[12] Gasteiger E, Hoogland C, Gattiker A, et al. 2005. The Proteomics Protocols Handbook-Chapter 52: Protein Identification and Analysis Tools on the ExPASy Server[M]. Humana Press, Totowa, pp. 571-607.
[13] Guo Y, Gan S. 2006. AtNAP, a NAC family transcription factor, has an important role in leaf senescence[J]. The Plant Journal, 46(4): 601-612.
[14] Ha C V, Nasr Esfahani M, Watanabe Y, et al. 2014. Genome- Wide Identification and Expression Analysis of the CaNAC Family Members in Chickpea during Development, Dehydration and ABA Treatments[J]. PLOS ONE, 9(12): e114107.
[15] He X, Mu R, Cao W, et al. 2005. AtNAC2, a transcription factor downstream of ethylene and auxin signaling pathways, is involved in salt stress response and lateral root development[J]. The Plant Journal, 44(6): 903-916.
[16] Hillocks R J, Minja E, Nahdy M S, et al. 2000. Diseases and pests of pigeonpea in eastern Africa: A review[J]. International Journal of Pest Management, 46(1): 7-18.
[17] Hu H, You J, Fang Y, et al. 2008. Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice[J]. Plant Molecular Biology, 67(1-2): 169-181.
[18] Hu R, Qi G, Kong Y, et al. 2010. Comprehensive analysis of NAC domain transcription factor gene family in Populus trichocarpa[J]. BMC Plant Biology, 10(1): 145.
[19] Jensen M K, Hagedorn P H, Torres-Zabala D, et al. 2008. Transcriptional regulation by an NAC (NAM-ATAF1, 2- CUC2) transcription factor attenuates ABA signalling for efficient basal defence towards Blumeria graminis f. sp. hordei in Arabidopsis[J]. The Plant Journal, 56(6): 867-880.
[20] Jensen M K, Rung J H, Gregersen P L, et al. 2007. The HvNAC6 transcription factor: A positive regulator of penetration resistance in barley and Arabidopsis[J]. Plant Molecular Biology, 65(1): 137-150.
[21] Jeong R D, Chandra-Shekara A, Kachroo A, et al. 2008. HRT- mediated hypersensitive response and resistance to Turnip crinkle virus in Arabidopsis does not require the function of TIP, the presumed guardee protein[J]. Molecular Plant-Microbe Interactions, 21(10): 1316-1324.
[22] Kim Y S, Kim S G, Park J E, et al. 2006. A membrane-bound NAC transcription factor regulates cell division in Arabi- dopsis[J]. The Plant Cell, 18(11): 3132-3144.
[23] Lu P L, Chen N Z, An R, et al. 2007. A novel drought-inducible gene, ATAF1, encodes a NAC family protein that negatively regulates the expression of stress-responsive genes in Arabidopsis[J]. Plant Molecular Biology, 63: 289-305.
[24] Oliveira M B, Junior M L, G-D-Sá M, et al. 2015. Exogenous application of methyl jasmonate induces a defense response and resistance against Sclerotinia sclerotiorum in dry bean plants[J]. Journal of Plant Physiology, 182: 13-22.
[25] Ooka H, Satoh K, Doi K, et al. 2003. Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana[J]. DNA Research, 10(6): 239-247.
[26] Pinheiro G L, Marques C S, Costa M D, et al. 2009. Complete inventory of soybean NAC transcription factors: Sequence conservation and expression analysis uncover their distinct roles in stress response[J]. Gene, 444(1-2): 10-23.
[27] Puranik S, Sahu P P, Srivastava P S, et al. 2012. NAC proteins: Regulation and role in stress tolerance[J]. Trends in Plant Science, 17(6): 369-381.
[28] Ren T, Qu F, Morris T J. 2000. HRT gene function requires interaction between a NAC protein and viral capsid protein to confer resistance to turnip crinkle virus[J]. The Plant Cell, 12(10): 1917-1925.
[29] Ren T, Qu F, Morris T J. 2005. The nuclear localization of the Arabidopsis transcription factor TIP is blocked by its interaction with the coat protein of Turnip crinkle virus[J]. Virology, 331(2): 316-324.
[30] Riechmann J L, Heard J, Martin G, et al. 2000. Arabidopsis transcription factors: Genome-wide comparative analysis among eukaryotes[J]. Science, 290(5499): 2105-2110.
[31] Satheesh V, Jagannadham P T K, Chidambaranathan P, et al. 2014. NAC transcription factor genes: genome-wide identification, phylogenetic, motif and cis-regulatory element analysis in pigeonpea (Cajanus cajan (L.) Millsp.)[J]. Molecular Biology Reports, 41(12): 7763-7773.
[32] Selth L A, Dogra S C, Rasheed M S, et al. 2005. A NAC domain protein interacts with tomato leaf curl virus replication accessory protein and enhances viral replication[J]. The Plant Cell, 17(1): 311-325.
[33] Singh S, Kudapa H, Garg V, et al. 2021. Comprehensive analysis and identification of drought responsive candidate NAC genes in three semi-arid tropics (SAT) legume crops[J]. BMC Genomics, 22: 289.
[34] Song Z H, Dong B Y, Yang Q, et al. 2020. Screening of CBL genes in pigeon pea with focus on the functional analysis of CBL4 in abiotic stress tolerance and flavonoid biosynthesis[J]. Environmental and Experimental Botany, 177: 104102.
[35] Sparkes I A, Runions J, Kearns A, et al. 2006. Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants[J]. Nature Protocols, 1(4): 2019-2025.
[36] Sun Q W, Huang J F, Guo Y, et al. 2020. A cotton NAC domain transcription factor, GhFSN5, negatively regulates secondary cell wall biosynthesis and anther development in transgenic Arabidopsis[J]. Plant Physiology and Biochemistry, 146: 303-314.
[37] Troedson R J, Wallis E S, Singh L. 1990. Pigeonpea: Adaptation[J]. The Pigeonpea, pp. 159-177.
[38] Uauy C, Distelfeld A, Fahima T, et al. 2006. A NAC Gene regulating senescence improves grain protein, zinc, and iron content in wheat[J]. Science, 314(5803): 1298-1301.
[39] Vroemen C W, Mordhorst A P, Albrecht C, et al. 2003. The CUP-SHAPED COTYLEDON3 gene is required for boundary and shoot meristem formation in Arabidopsis [J]. The Plant Cell, 15(7): 1563-1577.
[40] Wang X, Basnayake B M, Zhang H, et al. 2009. The Arabidop- sis ATAF1, a NAC transcription factor, is a negative regulator of defense responses against necrotrophic fungal and bacterial pathogens. Molecular Plant-Microbe Interactions, 22(10): 1227-1238.
[41] Wang X, Goregaoker S P, Culver J N.2009a. Interaction of the Tobacco mosaic virus replicase protein with a NAC domain transcription factor is associated with the suppression of systemic host defenses[J]. Journal of Virology, 83(19): 9720-9730.
[42] Wang Y, Bao Z, Zhu Y, et al. 2009b. Analysis of temperature modulation of plant defense against biotrophic microbes[J]. Molecular Plant-Microbe Interactions, 22(5): 498-506.
[43] Wilkins M R, Gasteiger E, Bairoch A, et al. 1999. Protein identification and analysis tools in the ExPASy server[J]. Methods in Molecular Biology, 112: 531-552.
[44] Willemsen V, Bauch M, Bennett T, et al. 2008. The NAC domain transcription factors FEZ and SOMBRERO control the orientation of cell division plane in Arabidopsis root stem cells[J]. Developmental Cell, 15(6): 913-922. Wu Y, Deng Z, Lai J, et al. 2009. Dual function of Arabidopsis ATAF1 in abiotic and biotic stress responses[J]. Cell Research, 19(11): 1279-1290.
[45] Xie Q, Sanz-Burgos A P, Guo H S, et al. 1999. GRAB proteins, novel members of the NAC domain family, isolated by their interaction with a geminivirus protein[J]. Plant Molecular Biology, 39(4): 647-656.
[46] Yoo S Y, Kim Y, Kim S Y, et al. 2007. Control of flowering time and cold response by a NAC-domain protein in Ara- bidopsis[J]. PLOS ONE, 2(7): e642.
[47] Yoshii M, Shimizu T, Yamazaki M, et al. 2009. Disruption of a novel gene for a NAC-domain protein in rice confers resistance to Rice dwarf virus[J]. The Plant Journal, 57(4): 615-625.
[48] Zheng X, Chen B, Lu G, et al. 2009. Overexpression of a NAC transcription factor enhances rice drought and salt tolerance[J]. Biochemical and Biophysical Research Communications, 379(4): 985-989.
[49] Zimmermann R, Werr W. 2005. Pattern formation in the monocot embryo as revealed by NAM and CUC3 orthologues from Zea mays L.[J]. Plant Molecular Biology, 58: 669-685. |
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