马铃薯StDof5的克隆及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2023-0960

摘要/Abstract

摘要：

【目的】 DNA binding with one finger（Dof）基因家族是植物特有的转录因子在植物生长发育和胁迫响应中发挥重要作用。研究Dof转录因子在马铃薯响应逆境胁迫特别是盐胁迫中的功能与机制，为获得抗盐逆境胁迫新品种提供一定的理论和技术基础。【方法】 克隆StDof5，并对其进行生物信息学分析，通过同源重组的方法构建StDof5过表达载体，转化马铃薯得到阳性植株，并对其表达进行分析。【结果】 StDof5的CDS全长498 bp，编码165个氨基酸，分子式为C₈₀₀H₁₂₃₆N₂₄₂O₂₄₆S₉，StDof5原子个数2 533个，相对分子量为18 468.63 kD，等电点是8.47，脂肪族指数为51.45，不稳定系数为45.16，预测该蛋白为不稳定蛋白。预测得到的跨膜螺旋个数为0，其属于非跨膜蛋白。StDof5蛋白亲水性平均值为-0.876，预测该蛋白属于亲水性蛋白。StDof5蛋白二级结构预测结果显示α螺旋占20%，β转角占0，延长链占9.7%，无规卷曲占66.06%。盐胁迫表型试验表明，过表达植株（OPs）比野生型（DES）的株高、鲜重均有明显程度上升，但根长、根数和叶片数差异不大。【结论】 过表达植株（OPs）能够抵御盐胁迫，对盐胁迫呈现脱敏感表型。该基因对盐胁迫响应明显，且存在组织特异性。

关键词: 马铃薯, StDof5, 载体构建, 遗传转化

Abstract:

【Objective】 The DNA binding with one finger（Dof）gene family is a plant-specific transcription factor family that plays important roles in plant growth, development and stress response. Studying the function and mechanism of Dof transcription factors responding to abiotic stress, especially salt stress in potato, provides some theoretical and technical basis for obtaining new varieties resistant to salt stress.【Method】 StDof5 was cloned, and it was analyzed via bioinformatics. StDof5 overexpression vector was constructed by homologous recombination, potato was transformed to obtain positive plants lines, and their expressions were analyzed.【Result】 CDS of StDof5 is 498 bp, it encodes 165 amino acids, and the molecular formula is C₈₀₀H₁₂₃₆N₂₄₂O₂₄₆S₉. The number of StDof5 atoms is 2 533, the relative molecular weight is 18 468.63 kD, the isoelectric point is 8.47, the aliphatic index is 51.45, and the instability coefficient is 45.16, the protein is predicted to be an unstable protein. The predicted number of StDof5 transmembrane helices is 0, thus it is a non-transmembrane protein. The average value of hydrophilicity of StDof5 protein is -0.876, predicting that it is hydrophilic. The predicted secondary structure of StDof5 protein showed it has 20% α-helices, 0 β-turn, 9.7% extended strands and 66.06% random coils. Salt stress phenotype experiments showed that the plant heights and fresh weights of overexpressing plants（OPs）were significantly higher than those of wild type（DES）plants, but there was no significant difference in root length, root number and leaf number between them. 【Conclusion】 Overexpressing plants（OPs）can resist salt stress and demonstrate desensitization phenotype to salt stress. StDof5 has significant responses to salt stress and tissue specificity.

Key words: potato, StDof5 gene, vector construction, genetic transformation

梅显军, 宋慧洋, 李京昊, 梅超, 宋倩娜, 冯瑞云, 陈喜明. 马铃薯StDof5的克隆及表达分析[J]. 生物技术通报, 2024, 40(3): 181-192.

MEI Xian-jun, SONG Hui-yang, LI Jing-hao, MEI Chao, SONG Qian-na, FENG Rui-yun, CHEN Xi-ming. Cloning and Expression Analysis of StDof5 Gene in Potato[J]. Biotechnology Bulletin, 2024, 40(3): 181-192.

图/表 12

表1 荧光定量引物序列

Table 1 Primer sequences for fluorescence quantification

引物名称Primer name	引物序列Primer sequence（5'-3'）
Actin-F	GGGATGGAGAAGTTTGGTGGTGG
Actin-R	CTTCGACCAAGGGATGGTGTAGC
Dof-F.2	GCTTGACTTTGATGGAGTGGTGG
Dof-R.2	AAGATTGGCTATCGGAGGTGC
ABF4-F	CTTCTGCACATACAAGGATATTCAA
ABF4-R	TGCCTAGCCAAAGGGAAATCA
MYB2-F	ATGACATGGACCTCCGACGA
MYB2-R	CGTTCGCTTTAGACCAGCAC
SnRK2s-F	TTGCTGTAGAGAAGGTGGGT
SnRK2s-R	GTCCTACTGTCACTGCCGTC
DREB2A-F	CACAGAAGAAAAGAGAAATGGCT
DREB2A-R	TCCACTATAATCCAACGGCAGA
ABI3-F	TGCTGACATGAAGTGTGGCA
ABI3-R	TTTTGCCCTCCGACTTTGGT
ABI5-F	TAAAACAGGCCCTGGCGGAG
ABI5-R	TTGCGCCTTTGTTTGAGCTT

图1 StDof5蛋白生物信息学分析 A：StDof5蛋白的跨膜结构域；B：马铃薯 StDof5蛋白的二级结构预测；C：马铃薯StDof5蛋白质的亲水性；D：马铃薯 StDof5蛋白的三级结构预测

Fig. 1 Bioinformatics analysis of StDof5 protein A: Transmembrane domain of the StDof5 protein; B: secondary structure prediction of potato StDof5 protein; C: hydrophilic nature of potato StDof5 protein; D: tertiary structure prediction of potato StDof5 protein

图2 StDof5氨基酸序列及其他植物同源序列间多序列比对

Fig. 2 Multiple sequence alignment between StDof5 amino acid sequences and other plants' homologous sequences

图3 不同植物中Dof5蛋白进化树

Fig. 3 Evolutionary tree of Dof5 proteins in different plants

图4 马铃薯StDof5的PCR扩增 1：含目的基因StDof5的质粒；2：Hind III-Sac I酶切质粒；M：1 kb marker

Fig. 4 PCR amplification of the potato StDof5 gene 1: Plasmid containing target gene StDof5. 2: Hind III-Sac I digested plasmid. M: 1 kb marker

图5 马铃薯StDof5载体的构建 A：农杆菌载体构建；B：过表达马铃薯植株鉴定；C：马铃薯遗传转化组织培养；M： marker 2000 bp；P：质粒；DES：野生型，1-8：阳性单克隆；6-10、14、16、24：过表达阳性植株

Fig. 5 Construction of the potato StDof5 vector A: Agrobacterium vector construction; B: potato genetically transformed tissue culture; C: identification of overexpressed potato plants. M: Marker 2000 bp. P: Plasmid. DES: Wild type. 1-8: Positive single clone. 6-10, 14, 16, 24: Overexpression positive plants

图6 不同组织中马铃薯Dof5的表达量分析 DES：野生型；OP：过表达阳性植株；不同字母表示差异显著（P<0.05）。下同

Fig. 6 Analysis of Dof5 gene expression in different tissues of potato DES: Wild type. OP: Overexpressed positive plants. Different letters indicate a significant difference（P<0.05）. The same below

图7 StDof5的表达量

Fig. 7 StDof5 gene expression

图8 过表达Dof5阳性植株中胁迫响应基因的表达量分析

Fig. 8 Expression analysis of stress-responsive genes in overexpressing Dof5-positive plants

表2 盐胁迫生长指标的测定

Table 2 Determination of the growth indicators of salt stress

测量指标Measurement index	盐浓度Salting strength/（mol·L^-1）	品种名称Variety name
测量指标Measurement index	盐浓度Salting strength/（mol·L^-1）	DES	OP-6	OP-7	OP-9	OP-10	OP-16	OP-24
株高Plant height/cm	0	6.87±0.38a	7.10±0.17a	5.57±0.40a	5.43±0.15a	6.53±0.85a	7.37±0.75a	7.83±1.01a
	50	3.70±010b	4.83±0.65b	3.70±0.36b	4.37±0.21b	3.97±0.35b	6.00±0.72b	4.13±0.45b
	100	2.13±0.76c	2.97±0.42c	2.00±0.53c	1.77±0.31c	2.80±0.36c	3.00±0.44c	2.93±0.15c
鲜重Fresh weight/g	0	0.21±0.01a	0.23±0.01a	0.19±0.01a	0.21±0.02a	0.24±0.02a	0.19±0.02a	0.35±0.01a
	50	0.14±0.01b	0.19±0.02b	0.12±0.02b	0.16±0.02b	0.16±0.02b	0.16±0.01a	0.28±0.02b
	100	0.09±0.01c	0.12±0.02c	0.09±0.01c	0.09±0.01c	0.12±0.01c	0.12±0.02b	0.15±0.02c
根长Root length/cm	0	12.13±0.25a	12.20±0.46a	12.23±2.03a	14.70±0.62a	17.63±0.35a	12.90±1.49a	12.03±0.72a
	50	9.00±0.26b	9.73±0.12b	10.93±1.79ab	13.07±0.31b	13.73±0.31b	10.53±0.61b	11.67±0.31a
	100	7.53±0.25c	8.93±060b	6.70±2.74b	4.00±0.72c	10.1±0.20c	9.73±0.31b	8.60±0.53b
根数Root	0	6.33±1.15a	7.33 ±2.31a	8.00±0.00a	7.00±1.00a	8.67±1.53a	6.00±0.00a	9.67±2.08a
	50	3.67±2.08a	5.67±2.31a	4.00±1.00b	5.33±2.31a	4.67±1.53b	6.67±2.89a	5.67±1.15b
	100	3.33±1.53a	5.00±1.00a	3.33±2.08b	1.00±0.00b	4.33±2.31b	1.67±0.58b	3.00±1.73b
叶片数Number of leaves	0	7.33±0.58a	8.33±0.58a	7.67±0.58a	6.67±0.58a	7.00±0.00a	7.00±1.00a	6.67±0.58a
	50	6.33±0.58a	7.00±0.00a	6.33±1.15a	6.00±1.00ab	7.00±0.00a	7.00±1.00a	6.66±0.58a
	100	7.00±1.00a	7.00±1.00a	6.67±0.58a	5.00±0.00b	6.33±1.15a	6.00±1.00a	6.67±0.58a

图9 盐胁迫25 d后组培苗生长状态图片从上到下依次是：DES、OP-6、OP-7、OP-9、OP-10、OP-16、OP-24，图片内分别为梯度0、50和100 mmol/L NaCl浓度的盐胁迫

Fig. 9 Growth status of tissue culture seedlings after 25 d of salt stress From top to bottom, the pictures are: DES, OP-6, OP-7, OP-9, OP-10, OP-16, OP-24, with the salt stress of 0, 50 and 100 mmol/L NaCl, respectively

图10 SOD、POD和CAT的酶活性

Fig. 10 Enzymatic activities of SOD, POD, and CAT

参考文献 48

[1]	Gupta S, Malviya N, Kushwaha H, et al. Insights into structural and functional diversity of Dof(DNA binding with one finger)transcription factor[J]. Planta, 2015, 241(3): 549-562. doi: 10.1007/s00425-014-2239-3 pmid: 25564353
[2]	Washio K. Functional dissections between GAMYB and Dof transcription factors suggest a role for protein-protein associations in the gibberellin-mediated expression of the RAmy1A gene in the rice aleurone[J]. Plant Physiol, 2003, 133(2): 850-863. pmid: 14500792
[3]	Wang CM, Song BB, Dai YQ, et al. Genome-wide identification and functional analysis of U-box E3 ubiquitin ligases gene family related to drought stress response in Chinese white pear(Pyrus bretschneideri)[J]. BMC Plant Biol, 2021, 21(1): 235. doi: 10.1186/s12870-021-03024-3
[4]	Wang T, Yue JJ, Wang XJ, et al. Genome-wide identification and characterization of the Dof gene family in moso bamboo(Phyllostachys heterocycla var. pubescens)[J]. Genes Genom, 2016, 38(8): 733-745. doi: 10.1007/s13258-016-0418-2 URL
[5]	Li Y, Ding WJ, Jiang HY, et al. Advances in research of the Dof gene family in plant[J]. Acta Botanica Boreali-Occidentalia Sinica, 2018, 38(9): 1758-1766.
[6]	Yanagisawa S, Schmidt RJ. Diversity and similarity among recognition sequences of Dof transcription factors[J]. Plant J, 1999, 17(2): 209-214. doi: 10.1046/j.1365-313x.1999.00363.x pmid: 10074718
[7]	Yanagisawa S. Dof domain proteins: Plant-specific transcription factors associated with diverse phenomena unique to plants[J]. Plant Cell Physiol, 2004, 45(4): 386-391. doi: 10.1093/pcp/pch055 pmid: 15111712
[8]	Venkatesh J, Park SW. Genome-wide analysis and expression profiling of DNA-binding with one zinc finger(Dof)transcription factor family in potato[J]. Plant Physiol Biochem, 2015, 94: 73-85. doi: 10.1016/j.plaphy.2015.05.010 URL
[9]	Yanagisawa S, Izui K. Molecular cloning of two DNA-binding proteins of maize that are structurally different but interact with the same sequence motif[J]. J Biol Chem, 1993, 268(21): 16028-16036. pmid: 8340424
[10]	张华珍, 吴昊, 李香花, 等. 一个低氮诱导表达的水稻Dof转录因子OsDof-13的分离和转化[J]. 分子植物育种, 2007, 5(4): 455-460.
	Zhang HZ, Wu H, Li XH, et al. Isolation and transformation of OsDof-13, a member of rice Dof transcription factor family, induced by low nitrogen stress[J]. Mol Plant Breed, 2007, 5(4): 455-460.
[11]	Lijavetzky D, Carbonero P, Vicente-Carbajosa J. Genome-wide comparative phylogenetic analysis of the rice and Arabidopsis Dof gene families[J]. BMC Evol Biol, 2003, 3: 17. pmid: 12877745
[12]	Cai XF, Zhang YY, Zhang CJ, et al. Genome-wide analysis of plant-specific Dof transcription factor family in tomato[J]. J Integr Plant Biol, 2013, 55(6): 552-566. doi: 10.1111/jipb.v55.6 URL
[13]	马凯恒, 张彤, 闫鹏宇, 等. 核桃转录因子JrDof3的克隆及渗透胁迫表达分析[J]. 中南林业科技大学学报, 2020, 40(10): 35-41.
	Ma KH, Zhang T, Yan PY, et al. Cloning and expression analysis of walnut transcription factor Dof3 to osmotic stress[J]. J Central South Univ For Technol, 2020, 40(10): 35-41.
[14]	Corrales AR, Nebauer SG, Carrillo L, et al. Characterization of tomato cycling Dof factors reveals conserved and new functions in the control of flowering time and abiotic stress responses[J]. J Exp Bot, 2014, 65(4): 995-1012. doi: 10.1093/jxb/ert451 URL
[15]	成亮, 刘宝玲, 刘盼, 等. 青狗尾草Dof基因家族的全基因组鉴定及组织特异性表达分析[J]. 分子植物育种, 2018, 16(13): 4226-4234.
	Cheng L, Liu BL, Liu P, et al. Genome-wide analysis and expression profiling of dof family in Setaria viridis[J]. Mol Plant Breed, 2018, 16(13): 4226-4234.
[16]	Choi HI, Hong JH, Ha JO, et al. ABFs, a family of ABA-responsive element binding factors[J]. J Biol Chem, 2000, 275(3): 1723-1730. doi: 10.1074/jbc.275.3.1723 pmid: 10636868
[17]	李杨. 玉米中PP2Cs与SnRK2s相互作用研究[D]. 合肥: 安徽农业大学, 2021.
	Li Y. Study on the interaction between PP2Cs and SnRK2s in maize[D]. Hefei: Anhui Agricultural University, 2021.
[18]	赵芝婧. 拟南芥转录因子LFY2和MYB2在花药发育调控网络中的作用研究[D]. 北京林业大学, 2020.
	Zhao ZJ. Role of Arabidopsis transcription factors LFY2 and MYB2 in anther development regulation network[D]. Beijing: Beijing Forestry University, 2020.
[19]	Achard P, Gong F, Cheminant S, et al. The cold-inducible CBF1 factor-dependent signaling pathway modulates the accumulation of the growth-repressing DELLA proteins via its effect on gibberellin metabolism[J]. Plant cell, 2008, 20(8): 2117-2129. doi: 10.1105/tpc.108.058941 pmid: 18757556
[20]	Fujita Y, Fujita AM, Satoh CR, et al. AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis[J]. Plant Cell, 2005, 17(12): 3470-3488. doi: 10.1105/tpc.105.035659 URL
[21]	Zheng ZF, Xu XP, Crosley RA, et al. The protein kinase SnRK2.6 mediates the regulation of sucrose metabolism and plant growth in Arabidopsis[J]. Plant Physiol, 2010, 153(1): 99-113. doi: 10.1104/pp.109.150789 URL
[22]	Wang YP, Li L, Ye TT, et al. The inhibitory effect of ABA on floral transition is mediated by ABI5 in Arabidopsis[J]. J Exp Bot, 2013, 64(2): 675-684. doi: 10.1093/jxb/ers361 URL
[23]	王伟, 李立芹, 邹雪, 等. 马铃薯块茎总RNA提取方法比较[J]. 江苏农业科学, 2013, 41(1): 38-40.
	Wang W, Li LQ, Zou X, et al. Comparison of extraction methods of total RNA from potato tuber[J]. Jiangsu Agric Sci, 2013, 41(1): 38-40.
[24]	于惠琳, 吴玉群, 王延波, 等. 甜玉米籽粒总RNA提取方法的比较[J]. 园艺与种苗, 2022, 42(1): 67-68, 71.
	Yu HL, Wu YQ, Wang YB, et al. Comparison of total RNA extraction methods from sweet corn kernels[J]. Hortic Seed, 2022, 42(1): 67-68, 71.
[25]	张晓丽, 代红军. 植物RNA提取方法的研究进展[J]. 北方园艺, 2014(8): 175-178.
	Zhang XL, Dai HJ. Research progress on extraction method of plant RNA[J]. North Hortic, 2014(8): 175-178.
[26]	赵勤. 农杆菌介导的马铃薯遗传转化影响因素研究进展[J]. 园艺与种苗, 2011, 31(2): 92-94.
	Zhao Q. Study on influencing factors of Agrobacterium-mediated genetic transformation in potato[J]. Rain Fed Crops, 2011, 31(2): 92-94.
[27]	杨永智. 农杆菌介导法马铃薯遗传转化体系的优化[J]. 江苏农业学报, 2013, 29(4): 738-742.
	Yang YZ. Optimization of Agrobacterium tumerfaciens mediated potato transformation system[J]. Jiangsu J Agric Sci, 2013, 29(4): 738-742.
[28]	连欢欢, 杨永智, 王舰. 根癌农杆菌介导的马铃薯DM茎段遗传转化条件的优化[J]. 分子植物育种, 2016, 14(6): 1491-1499.
	Lian HH, Yang YZ, Wang J. Optimization of conditions for genetic transformation of potato DM stems[J]. Molecular plant breeding, 2016, 14(6): 1491-1499.
[29]	张萍. 根癌农杆菌介导的马铃薯遗传转化体系的研究[J]. 陕西农业科学, 2019, 65(4): 27-29, 40.
	Zhang P. Study on genetic transformation system of potato mediated by Agrobacterium tumefaciens[J]. Shaanxi J Agric Sci, 2019, 65(4): 27-29, 40.
[30]	李逸菲, 王瑞博, 夏惠婷, 等. 农杆菌介导四季秋海棠遗传转化体系建立[J]. 东北农业大学学报, 2023, 54(5): 19-27.
	Li YF, Wang RB, Xia HT, et al. Establishment of Agrobacteri-um-mediated genetic transformation system for Begonia semperflo-rens[J]. J Northeast Agric Univ, 2023, 54(5): 19-27.
[31]	宋倩娜, 梅超, 霍利光, 等. 马铃薯品种‘并薯6号’遗传转化体系的建立[J]. 中国马铃薯, 2021, 35(5): 385-396.
	Song QN, Mei C, Huo LG, et al. Establishment of the genetic transformation system for potato variety ‘Bingshu 6’[J]. Chin potato J, 2021, 35(5): 385-396.
[32]	高志鹏, 杜玉萧, 冯霞, 等. 鄂马铃薯3号遗传转化体系的构建与表达[J]. 湖北农业科学, 2022, 61(9): 162-167, 191.
	Gao ZP, Du YX, Feng X, et al. Construction and expression of the # 3 genetic transformation system of Hubei potato No.3[J]. Hubei Agric Sci, 2022, 61(9): 162-167, 191.
[33]	陈兆贵, 林燕文, 李希陶, 等. 马铃薯植株再生和遗传转化技术研究进展[J]. 智慧农业导刊, 2023, 3(9): 12-15.
	Chen ZG, Lin YW, Li XT, et al. Research progress of plant regeneration and genetic transformation of potatoes[J]. J Smart Agri, 2023, 3(9): 12-15.
[34]	公鑫, 王业红, 张剑峰, 等. 马铃薯RPP13基因RNAi载体的构建及遗传转化[J]. 华北农学报, 2023, 38(1): 178-183. doi: 10.7668/hbnxb.20193453
	Gong X, Wang YH, Zhang JF, et al. Construction and genetic transformation of the RNAi vector of potato RPP 13 gene[J]. Acta Agric Boreali Sin, 2023, 38(1): 178-183.
[35]	万丽丽, 王转茸, 汤谧, 等. 甜瓜遗传转化技术的优化及DRs类基因的应用[J]. 江苏农业科学, 2023, 51(7): 49-58.
	Wan LL, Wang ZR, Tang M, et al. Optimization of genetic transformation technology of melon and the application of DRs class genes[J]. Jiangsu Agric Sci, 2023, 51(7): 49-58.
[36]	Corrales AR, Carrillo L, Lasierra P, et al. Multifaceted role of cycling DOF factor 3(CDF3)in the regulation of flowering time and abiotic stress responses in Arabidopsis[J]. Plant Cell Environ, 2017, 40(5): 748-764. doi: 10.1111/pce.v40.5 URL
[37]	Xu PP, Chen HY, Ying L, et al. AtDOF5.4/OBP4, a DOF transcription factor gene that negatively regulates cell cycle progression and cell expansion in Arabidopsis thaliana[J]. Sci Rep, 2016, 6: 27705. doi: 10.1038/srep27705
[38]	van Zelm E, Zhang YX, Testerink C. Salt tolerance mechanisms of plants[J]. Annu Rev Plant Bio, 2020, 71: 403-433. doi: 10.1146/annurev-arplant-050718-100005 URL
[39]	Su Y, Liang W, Liu ZJ, et al. Overexpression of GhDof1 improved salt and cold tolerance and seed oil content in Gossypium hirsutum[J]. J Plant Physiol, 2017, 218: 222-234. doi: 10.1016/j.jplph.2017.07.017 URL
[40]	Cai XF, Zhang CJ, Shu WB, et al. The transcription factor SlDof22 involved in ascorbate accumulation and salinity stress in tomato[J]. Biochem Biophys Res Commun, 2016, 474(4): 736-741. doi: 10.1016/j.bbrc.2016.04.148 URL
[41]	Kim J, Kim JH, Lyu JI, et al. New insights into the regulation of leaf senescence in Arabidopsis[J]. J Exp Bot, 2018, 69(4): 787-799. doi: 10.1093/jxb/erx287 URL
[42]	Lee BD, Ri KM, Young KM, et al. The F-box protein FKF1inhibits dimerization of COP1 in the control of photoperiodic flowering[J]. Nature Communications, 2018, 8(1): 2259. doi: 10.1038/s41467-017-02476-2
[43]	Wen CL, Cheng Q, Zhao LQ, et al. Identification and characterisation of Dof transcription factors in the cucumber genome[J]. Sci Rep, 2016, 6: 23072. doi: 10.1038/srep23072
[44]	Noguero M, Atif RM, OCHATT S, et al. The role of the DNA-binding One Zinc Finger(Dof)transcription factor family in plants[J]. Plant Sci, 2013, 209: 32-45. doi: 10.1016/j.plantsci.2013.03.016 URL
[45]	Zhang ZR, Yuan L, Liu X, et al. Evolution analysis of Dof transcription factor family and their expression in response to multiple abiotic stresses in Malus domestica[J]. Gene, 2018, 639: 137-148. doi: 10.1016/j.gene.2017.09.039 URL
[46]	Ewas M, Khames E, et al. The tomato Dof Daily Fluctuations 1, DESDF1 acts as flowering accelerator and protector against various stresses[J]. Sci Rep, 2017, 7(1): 10299. doi: 10.1038/s41598-017-10399-7
[47]	Ma J, Li MY, Wang F, et al. Genome-wide analysis of Dof family transcription factors and their responses to abiotic stresses in Chinese cabbage[J]. BMC Genomics, 2015, 16(1): 33. doi: 10.1186/s12864-015-1242-9 URL
[48]	Kang Y, Yang X, Liu Y, et al. Integration of mRNA and miRNA analysis reveals the molecular mechanism of potato(Solanum tuberosum L.)response to alkali stress[J]. International Journal of Biological Macromolecules, 2021, 182: 938-949. doi: 10.1016/j.ijbiomac.2021.04.094 URL