Abstract:Industrial hemp (Cannabis sativa) is an important economic crop, and its seed germination period is susceptible to drought stress, which ultimately has adverse effects on the growth and yield of hemp. NAC gene is a unique type of transcription factor in plants, which plays an essential role in plant response to abiotic stress. Based on the previous transcriptome data, a NAC gene from industrial hemp 'Yunma 1' was cloned by RT-PCR in this study, and named as CsNAC62 (GenBank No. XM_030652694). The results of sequence analysis found that the coding sequence of CsNAC62 gene contains 1 260 bp, encoding 419 amino acids, its protein molecular weight is 48.32 kD, and the isoelectric point is 8.24. The N-terminal of CsNAC62 contains a NAC domain composed of 150~160 amino acids. This domain is composed of 5 subdomains, which is a typical conservative domain of the NAM/NAC gene family. Phylogenetic analysis showed that CsNAC62 was closely related to the NAC3 protein of peach (Prunus persica) in all the tested species. The gene expression level of CsNAC62 in the germinated seeds of industrial hemp 'Yunma 1' gradually increased with the prolongation of the drought stress treatment (PEG-6000 simulation), and the expression level reached to the maximumon the 7 d of germinated seeds. In normal seed germination (control), the expression level of CsNAC62 increased slowly. These results indicated that the gene expression of CsNAC62 was induced by drought. This study preliminarily explored the expression mode of the CsNAC62 under PEG simulated drought stress, and provides a reference for further exploring its molecular mechanism of participating in the response to industrial cannabis drought stress.
[1] 胡华冉. 2019. 工业大麻耐盐候选基因GDH2和NACs的克隆及功能分析[D]. 博士学位论文,云南大学, 导师: 刘飞虎, pp. 110-111. (Hu H R, 2019. Cloning and functional analysis of potential salt-tolerant genes of GDH2 and NACs in industrial hemp (Cannabis sativa L.)[D]. Thesis for Ph.D., Yunnan University, Suppervisor: Liu F H, pp. 110-111.) [2] 靳进朴, 郭安源, 何坤, 等. 2015. 植物转录因子分类、预测和数据库构建[J]. 生物技术通报, 31(11): 68-77. (Jin J P, Guo A Y, He K, et al.2015. Classification, prediction and database construction of plant transcription factors[J]. Biotechnology Bulletin, 31(11): 68-77.) [3] 卢延旭, 董鹏, 崔晓光, 等. 2007. 工业大麻与毒品大麻的区别及其可利用价值[J]. 中国药理学通报, 23(8): 1112-1114. (Lu Y X, Dong P, Cui X G, et al.2007. An approach for the analysis of pharmacodynamic interactions and the simulation of combined response[J]. Chinese Pharmacological Bulletin, 23(8): 1112-1114.) [4] 马雪祺, 阴艳红, 冯婧娴, 等. 2021. 植物NAC转录因子研究进展[J]. 植物生理学报, 57(12): 2225-2234. (Ma X Q, Yin Y H, Feng J X, et al.2021. Research progress of NAC transcription factors in plant[J]. Plant Physiology Journal, 57(12): 2225-2234.) [5] 石汝杰, 胡廷章. 2009. 渗透胁迫对4个玉米品种种子萌发及幼苗生长的影响[J]. 种子, 28(7): 85-87. (Shi R J, Hu T Z.2009. Effects of permeating stress on seeds germination and seedlings growth of four maize varieties[J]. Seed, 28(7): 85-87.) [6] 熊和平. 2008. 麻类作物育种学[M]. 中国农业科学技术出版社, 北京. pp. 208-296. (Xiong H P.2008. Hemp Crop Breeding [M]. China Agricultural Science and Technology Press, Beijing. pp. 208-296.) [7] 赵越, 王晓楠, 孙宇峰, 等. 2021. 工业大麻纤维产量、品质影响因素及纤维发育相关基因研究进展[J]. 中国麻业科学, 43(3):155-160. (Zhao Y, Wang X N, Sun Y F, et al.2021. Factors influencing yield and quality of hemp fiber and genes potentially related to fiber development[J]. Plant Fiber Sciences in China, 43(3): 155-160.) [8] Aida M, Ishida T, Fukaki H, et al.1997. Genes involved in organ separation in Arabidopsis: An analysis of the cup-shaped cotyledon mutant[J]. The Plant Cell, 9(6): 841-857. [9] Boehnke K F, Gagnier J J, Matallana L, et al.2021. Substituting cannabidiol for opioids and pain medications among individuals with fibromyalgia: A large online survey[J]. The Journal of Pain, 22(11): 1418-1428. [10] Cai S, Zhang Z, Huang S, et al.2021. CannabisGDB: A comprehensive genomic database for Cannabis sativa L.[J]. Plant Biotechnology Journal, 19(5): 857-859. [11] Ding N, Zhao Y, Wang W, et al.2023. Transcriptome analysis in contrasting maize inbred lines and functional analysis of five maize NAC genes under drought stress treatment[J]. Frontiers in Plant Science, 13: 1097719. [12] Dudhate A, Shinde H, Yu P, et al.2021. Comprehensive analysis of NAC transcription factor family uncovers drought and salinity stress response in pearl millet (Pennisetum glaucum)[J]. BMC Genomics, 22(1): 70. [13] Geng L, Su L, Fu L, et al.2022. Genome-wide analysis of the rose (Rosa chinensis) NAC family and characterization of RcNAC091[J]. Plant Molecular Biology, 108(6): 1-15. [14] Li X, Wang Q, Guo C, et al.2022a. NtNAC053, a novel NAC transcription factor, confers drought and salt tolerances in tobacco[J]. Frontiers in Plant Science, 13: 817106. [15] Li Y, Zhang T, Xing W, et al.2022b. Comprehensive genomic characterization of the NAC transcription factors and their response to drought stress in Dendrobium catenatum[J]. Agronomy, 12(11): 2753. [16] Saidi M N, Mergby D, Souibgui A, et al.2022. Overexpression of durum wheat NAC transcription factor TtNTL3A promotes early flowering and increases multiple stress tolerance in transgenic Arabidopsis[J]. Plant Physiology and Biochemistry, 192: 1-9. [17] Srivastava R, Kobayashi Y, Koyama H, et al.2022. Cowpea NAC1/NAC2 transcription factors improve growth and tolerance to drought and heat in transgenic cowpea through combined activation of photosynthetic and antioxidant mechanisms[J]. Journal of Integrative Plant Biology, 60(1): 25-44. [18] Wang X, Basnayake B M, Zhang H, et al.2009. The Arabidopsis ATAF1, a NAC transcription factor, is a negative regulator of defense responses against necrotrophic fungal and bacterial pathogens[J]. Molecular Plant-Microbe Interactions, 22(10): 1227-1238. [19] Yang C, Huang Y, Lv P, et al.2022. NAC transcription factor GmNAC12 improved drought stress tolerance in soybean[J]. International Journal of Molecular Sciences, 23(19): 12029. [20] Yuan X, Wang H, Cai J, et al.2019. Rice NAC transcription factor ONAC066 functions as a positive regulator of drought and oxidative stress response[J]. BMC Plant Biology, 19(1): 278. [21] Zhao X L, Wei X Y, Guo Y, et al.2022. Industrial hemp-an old but versatile bast fiber crop[J]. Journal of Natural Fibers, 19(13): 6269-6282.