基于月季扩展蛋白基因家族鉴定的野蔷薇RmEXPB2基因功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2025-0216

摘要/Abstract

摘要：

目的系统鉴定中国古老月季‘月月粉’（Rosa chinensis ‘Old Blush’）扩展蛋白（Expansin）基因家族，解析其特征与表达模式，挖掘皮刺发育相关基因，并基于该结果进一步探究野蔷薇（Rosa multiflora）RmEXPB2基因的生物学功能，为深入鉴定扩展蛋白基因在蔷薇属植物皮刺形成中的功能奠定基础。方法利用生物信息学鉴定‘月月粉’基因组中的RcEXP，对其编码蛋白的氨基酸数量范围以及在染色体上的分布状况进行分析。通过构建系统进化树，剖析拟南芥（Arabidopsis thaliana）、‘月月粉’月季和森林草莓（Fragari avesca）的扩展蛋白（AtEXPs、RcEXPs和FvEXPs）的进化关系。检测‘月月粉’不同组织RcEXP表达模式及皮刺转录组中RcEXP表达。以皮刺研究模式植物野蔷薇作为实验材料，分离出候选基因RmEXPB2，研究该基因在皮刺发育进程中的表达变化情况，通过拟南芥异源表达探索其生物学功能。结果在‘月月粉’月季基因组中共鉴定出29个RcEXP，其编码蛋白含243‒313个氨基酸，在7条染色体上不均匀分布。系统进化树分析将AtEXPs、RcEXPs和FvEXPs分为4个亚组。在不同组织器官中，15个RcEXP表达模式各异，其中RcEXPB2和RcEXPA4在皮刺组织中优势表达。皮刺转录组数据中筛选获得13个RcEXP，在皮刺发育的特定阶段显著表达。从野蔷薇中分离出候选基因RmEXPB2，其在皮刺中优势表达，并且在皮刺发育末期表达量最高。异源表达RmEXPB2显著促进转基因拟南芥叶片扩展与根毛生长。结论月季‘月月粉’基因组中共鉴定出29个RcEXPs，其中13个成员在皮刺发育的特定阶段显著表达。野蔷薇RmEXPB2是一个皮刺优势表达基因，可能在皮刺伸长过程中发挥积极作用。

关键词: ‘月月粉’月季, 扩展蛋白, 野蔷薇, RmEXPB2, 皮刺发育, 异源表达, 系统进化分析, 组织表达模式

Abstract:

Objective This study is aimed to systematically identify the expansin gene family in Rosa chinensis ‘Old Blush’, dissect their sequence characteristics and expression patterns, screen genes associated with prickle development. Based on this result, the biological function of RmEXPB2 gene in Rosa multiflora was further explored in order to lay the foundation for in-depth identification of the function of expansin genes in prickle formation in Rosa sp. plants. Method Bioinformatics tools were used to identify RcEXP genes in R. chinensis ‘Old Blush’ genome. The amino acid residue range of their encoded proteins and chromosomal distribution were analyzed. A phylogenetic tree was constructed to analyze the evolutionary relationships among expansins from Arabidopsis thaliana (AtEXPs), R. chinensis ‘Old Blush’(RcEXPs), and Fragaria vesca (FvEXPs). RcEXP expression patterns in different tissues were detected, and its expression in the prickle transcriptome was analyzed. R. multiflora, a model for prickle research, served as the experimental material, the candidate gene RmEXPB2 was isolated, and its expression pattern during prickle development were studied. The biological function of RmEXPB2 was preliminarily explored by heterologous expression in Arabidopsis. Result A total of 29 RcEXP genes were identified in the genome of R. chinensis ‘Old Blush’. The encoded proteins ranged from 243 to 313 amino acids, and they were unevenly distributed on 7 chromosomes. Phylogenetic tree analysis divided the expansins AtEXPs, RcEXPs, and FvEXPs into 4 subgroups. Among different tissues and organs, 15 RcEXP genes showed diverse expression patterns, with RcEXPB2 and RcEXPA4 predominantly expressed in prickles. The 13 RcEXP genes were screened from the transcriptome data of prickles, which were significantly expressed at specific stages of prickle development. The gene RmEXPB2, preferentially expressed in the prickles, was isolated from R. multiflora, and its expression increased during prickle development, peaking at the late stage. Heterologous expression of RmEXPB2 significantly promoted leaf expansion and root hair growth in transgenic Arabidopsis. Conclusion A total of 29 RcEXPs were identified in the genome of R. chinensis ‘Old Blush’, of which 13 members were significantly expressed at specific stages of prickle development. RmEXPB2 is a prickle preferentially expressed gene in R. multiflora, which may play an active role in the process of prickle elongation.

Key words: Rosa chinensis ‘Old Blush’, expansin, Rosa multiflora, RmEXPB2, prickle development, heterologous expression, phylogenetic analysis, tissue expression pattern

李玉珍, 李梦丹, 张蔚, 彭婷. 基于月季扩展蛋白基因家族鉴定的野蔷薇RmEXPB2基因功能研究[J]. 生物技术通报, 2025, 41(9): 182-194.

LI Yu-zhen, LI Meng-dan, ZHANG Wei, PENG Ting. Functional Study of RmEXPB2 Genein Rosa multiflora Based on the Identification of the Expansin Gene Family in Rosa sp.[J]. Biotechnology Bulletin, 2025, 41(9): 182-194.

图/表 11

表1 RT-qPCR引物序列

Table 1 Primer sequences for RT-qPCR

Gene name	Forward primer （5′‒3′）	Reverse primer （5′‒3′）
RmUBC	GCCAGAGATTGCCCATATGTA	TCACAGAGTCCTAGCAGCACA
AtEF1α	GCAAGATGGATGCCACTACCC	GCAAGATGGATGCCACTACCC
RmEXPB2	TCTAGAATGGCTAGTTTAAGAAGCAT	CCCGGGTAGATAATTGACCACGGATC
RmEXPB2-qRT	TGCTGCTTGTTTGAGCAGAG	AAGCTCCACATTCCCTGCC
RmEXPB2-eGFP	GAGAACACGGGGGACTCTAGAATGGCTAGTTTAAGAAGCAT	GCCCTTGCTCACCATCCCGGGTAGATAATTGACCACGGATC

表2 ‘月月粉’月季扩展蛋白基因家族成员基本信息

Table 2 Basic information of expansin gene family members in R. chinensis'Old Blush'

基因ID Gene ID	基因 Gene	染色体位置 Chr location	等电点 pI	分子量 Molecular weight （Da）	蛋白长度 Protein length （aa）	亚细胞定位Subcellular localization	信号肽长度Signal peptie length （aa）
RchiOBHmChr5g0072761	RcEXPA1	Chr5	8.12	27 145.26	254	Cell wall	26
RchiOBHmChr7g0178811	RcEXPA2	Chr7	8.82	28 302.47	256	Cell wall	22
RchiOBHmChr7g0199891	RcEXPA3	Chr7	9.55	29 064.3	262	Chloroplast	28
RchiOBHmChr7g0181621	RcEXPA4	Chr7	8.64	26 524.79	250	Cell wall	20
RchiOBHmChr6g0281211	RcEXPA5	Chr6	6.93	27 816.75	253	Cell wall	23
RchiOBHmChr3g0488771	RcEXPA6	Chr3	7.55	29 302.32	272	Cell wall	24
RchiOBHmChr3g0470931	RcEXPA7	Chr3	9.00	28 950.9	260	Chloroplast	24
RchiOBHmChr4g0439541	RcEXPA8	Chr4	9.56	31 940.33	297	Mitochondrion
RchiOBHmChr3g0470941	RcEXPA9	Chr3	5.68	29 434.08	266	Cell wall	24
RchiOBHmChr5g0033911	RcEXPA10	Chr5	9.69	27 795.93	255	Cell wall	30
RchiOBHmChr1g0372131	RcEXPA11	Chr1	7.99	26 919.1	254	Cell wall	25
RchiOBHmChr3g0470961	RcEXPA12	Chr3	9.68	31 679.04	286	Chloroplast	25
RchiOBHmChr6g0245581	RcEXPA13	Chr6	5.22	28 332.98	260	Cell wall	27
RchiOBHmChr1g0367791	RcEXPA14	Chr1	9.53	27 917.86	259	Chloroplast	22
RchiOBHmChr3g0466461	RcEXPA15	Chr3	9.50	27 787.56	259	Cell wall	21
RchiOBHmChr2g0166091	RcEXPA16	Chr2	8.10	27 554.38	250	Chloroplast	22
RchiOBHmChr5g0065301	RcEXPA17	Chr5	9.68	27 890.68	260	Cell wall	22
RchiOBHmChr3g0460691	RcEXPA18	Chr3	8.36	27 487.08	255	Cell wall	24
RchiOBHmChr6g0303551	RcEXPA19	Chr6	8.87	27 306.82	257	Chloroplast	23
RchiOBHmChr2g0140121	RcEXPA20	Chr2	8.39	35 186.7	313	Cell wall	21
RchiOBHmChr2g0167701	RcEXPA21	Chr2	9.35	26 293.59	243	Cell wall	20
RchiOBHmChr2g0166111	RcEXPA22	Chr2	6.06	27 581.6	253	Cell wall	23
RchiOBHmChr5g0084081	RcEXPA23	Chr5	8.85	27 781.15	261	Nucleus	22
RchiOBHmChr2g0126231	RcEXPB1	Chr2	8.73	27 394.3	255	Cell wall	23
RchiOBHmChr1g0373921	RcEXPB2	Chr1	4.97	28 762.08	273	Chloroplast	23
RchiOBHmChr1g0348851	RcEXPB3	Chr1	6.40	30 068.99	287	Chloroplast	29
RchiOBHmChr6g0279781	RcEXLA1	Chr6	8.8	30 176.7	278	Chloroplast
RchiOBHmChr7g0205401	RcEXLB1	Chr7	6.87	27 333.67	250	Cell wall	22
RchiOBHmChr7g0208241	RcEXLB2	Chr7	4.70	28 099.57	258	Chloroplast	25

表2 ‘月月粉’月季扩展蛋白基因家族成员基本信息

Table 2 Basic information of expansin gene family members in R. chinensis'Old Blush'

基因ID Gene ID	基因 Gene	染色体位置 Chr location	等电点 pI	分子量 Molecular weight （Da）	蛋白长度 Protein length （aa）	亚细胞定位Subcellular localization	信号肽长度Signal peptie length （aa）
RchiOBHmChr5g0072761	RcEXPA1	Chr5	8.12	27 145.26	254	Cell wall	26
RchiOBHmChr7g0178811	RcEXPA2	Chr7	8.82	28 302.47	256	Cell wall	22
RchiOBHmChr7g0199891	RcEXPA3	Chr7	9.55	29 064.3	262	Chloroplast	28
RchiOBHmChr7g0181621	RcEXPA4	Chr7	8.64	26 524.79	250	Cell wall	20
RchiOBHmChr6g0281211	RcEXPA5	Chr6	6.93	27 816.75	253	Cell wall	23
RchiOBHmChr3g0488771	RcEXPA6	Chr3	7.55	29 302.32	272	Cell wall	24
RchiOBHmChr3g0470931	RcEXPA7	Chr3	9.00	28 950.9	260	Chloroplast	24
RchiOBHmChr4g0439541	RcEXPA8	Chr4	9.56	31 940.33	297	Mitochondrion
RchiOBHmChr3g0470941	RcEXPA9	Chr3	5.68	29 434.08	266	Cell wall	24
RchiOBHmChr5g0033911	RcEXPA10	Chr5	9.69	27 795.93	255	Cell wall	30
RchiOBHmChr1g0372131	RcEXPA11	Chr1	7.99	26 919.1	254	Cell wall	25
RchiOBHmChr3g0470961	RcEXPA12	Chr3	9.68	31 679.04	286	Chloroplast	25
RchiOBHmChr6g0245581	RcEXPA13	Chr6	5.22	28 332.98	260	Cell wall	27
RchiOBHmChr1g0367791	RcEXPA14	Chr1	9.53	27 917.86	259	Chloroplast	22
RchiOBHmChr3g0466461	RcEXPA15	Chr3	9.50	27 787.56	259	Cell wall	21
RchiOBHmChr2g0166091	RcEXPA16	Chr2	8.10	27 554.38	250	Chloroplast	22
RchiOBHmChr5g0065301	RcEXPA17	Chr5	9.68	27 890.68	260	Cell wall	22
RchiOBHmChr3g0460691	RcEXPA18	Chr3	8.36	27 487.08	255	Cell wall	24
RchiOBHmChr6g0303551	RcEXPA19	Chr6	8.87	27 306.82	257	Chloroplast	23
RchiOBHmChr2g0140121	RcEXPA20	Chr2	8.39	35 186.7	313	Cell wall	21
RchiOBHmChr2g0167701	RcEXPA21	Chr2	9.35	26 293.59	243	Cell wall	20
RchiOBHmChr2g0166111	RcEXPA22	Chr2	6.06	27 581.6	253	Cell wall	23
RchiOBHmChr5g0084081	RcEXPA23	Chr5	8.85	27 781.15	261	Nucleus	22
RchiOBHmChr2g0126231	RcEXPB1	Chr2	8.73	27 394.3	255	Cell wall	23
RchiOBHmChr1g0373921	RcEXPB2	Chr1	4.97	28 762.08	273	Chloroplast	23
RchiOBHmChr1g0348851	RcEXPB3	Chr1	6.40	30 068.99	287	Chloroplast	29
RchiOBHmChr6g0279781	RcEXLA1	Chr6	8.8	30 176.7	278	Chloroplast
RchiOBHmChr7g0205401	RcEXLB1	Chr7	6.87	27 333.67	250	Cell wall	22
RchiOBHmChr7g0208241	RcEXLB2	Chr7	4.70	28 099.57	258	Chloroplast	25

图1 ‘月月粉’月季expansin基因家族成员的染色体定位

Fig. 1 Chromosomal locations of expansin genefamily members in R. chinensis'Old Blush'

图2 ‘月月粉’月季、森林草莓和拟南芥的expansin基因家族进化树红色五角星代表‘月月粉’月季，蓝色圆形代表森林草莓，绿色方框代表拟南芥

Fig. 2 Phylogenetic tree of expansin gene family in R. chinensis'Old Blush', F. avesca and ArabidopsisThe red five-pointed star indicates R.chinensis 'Old Blush', the blue circle indicates F. avesca, and the green box indicates Arabidopsis

图3 ‘月月粉’月季expansin基因家族保守基序及基因结构分析A：保守基序分析；B：保守结构域分析；C：基因结构分析

Fig. 3 Analysis of conserved motif and gene structure of expansin gene family of R. chinensis'Old Blush'A: Conservative motif analysis. B: Conserved domain analysis. C: Gene structure analysis

图4 ‘月月粉’月季expansin基因启动子顺式元件分析

Fig. 4 Analysis of the cis-acting elements on the promoters of expansin gene in R. chinensis'Old Blush'

图5 ‘月月粉’月季部分expansin基因的组织表达分析

Fig. 5 Expression pattern analysis of some expansin gene in various tissues of R. chinensis'Old Blush'

图6 ‘月月粉’月季部分expansin基因在不同发育阶段皮刺中的表达分析（A）及基因数量（B）Stage I：柔软幼嫩、尚未木质化的皮刺；Stage Ⅱ：一定程度木质化、尖端开始向基部弯曲的皮刺；Stage Ⅲ：木质化较完全、形态基本成熟的皮刺；Stage IV：发育成熟、并初步开始衰老的皮刺

Fig. 6 Expression pattern analysis (A) and the gene number (B) of some expansin genes in different developmental stages of prickles in R. chinensis'Old Blush'Stage I: Soft and tender prickles that have not yet lignified. Stage Ⅱ: Prickles that are lignified to a certain extent and whose tips begin to bend toward the base. Stage Ⅲ: Prickles with relatively complete lignification and basically mature morphology. Stage IV: Prickles that have matured and initially begun to age

图7 RmEXPB2的表达量分析A：RmEXPB2在野蔷薇不同组织中的表达水平；B：RmEXPB2在野蔷薇不同发育阶段皮刺组织中的表达水平；C：野生型及T1代转基因株系RmEXPB2表达量检测；D：野生型及T2代转基因株系RmEXPB2表达量检测；Stage I：野蔷薇起始期皮刺；Stage Ⅱ：快速生长期皮刺；Stage Ⅲ：过渡期皮刺；Stage IV：硬化期皮刺；Stage V成熟期皮刺；line 1‒10代表转基因株系1‒10，WT代表野生型。ns表示P>0.05；*P<0.05；**P<0.01；***P<0.001。下同

Fig. 7 Expression analysis of RmEXPB2A: Expressions of RmEXPB2 in different tissues of R. multiflora. B: Expressions of RmEXPB2 in the prickles at different developmental stages. C: RmEXPB2 expression levels in WT and RmEXPB2-OE (T1) Arabidopsis. D: RmEXPB2 expression levels in WT and RmEXPB2-OE (T2) Arabidopsis. Stage I: Starting period of R. multiflora prickles; Stage II: rapid growth period prickles; Stage III: transition period prickles; Stage IV: hardening period prickles; Stage V: maturation period prickles; line 1-10 indicates transgenic line 1‒10, WT indicates wild type. ns indicates P>0.05; * indicates P<0.05; ** indicates P<0.01; *** indicates P<0.001. The same below

图8 RmEXPB2转基因拟南芥T2代株系的叶片表型A：莲座叶表型图；B：莲座叶数量，n>20；C：第1、3、5、6、8片莲座叶的叶长、叶宽、叶面积；Line 1、5、6代表转基因株系1、5、6，WT代表野生型。下同

Fig. 8 Leaf phenotypes of RmEXPB2-OE (T2) Arabidopsis plantsA: True leaf phenotypic diagram. B: Number of true leaves. n>20. C: The 1st, 3rd, 5th, 6th, 8th leaf length, width and area of the true leaf. Line1, 5, 6 indicates transgenic line1, 5, 6, WT indicates wild type. The same below

图9 RmEXPB2转基因拟南芥T2代植株的根部表型A：野生型及RmEXPB2转基因拟南芥的根；B：野生型及RmEXPB2转基因拟南芥的根毛；C‒D：野生型与RmEXPB2转基因拟南芥的主根长度、下胚轴长度；E‒F：野生型及RmEXPB2转基因拟南芥的根毛长度、根毛密度；n>20

Fig. 9 Root phenotypic analysis of RmEXPB2-OE (T2) Arabidopsis plantsA: Roots of WT and RmEXPB2-OE (T2) transgenic Arabidopsis. B: Root hairs of WT and RmEXPB2-OE (T2) Arabidopsis. C‒D: The main root length and hypocotyl length of WT and RmEXPB2-OE (T2) transgenic Arabidopsis. E‒F: The root hair length and root hair density of RmEXPB2-OE (T2) Arabidopsis. n>20

参考文献 36

[1]	Nakamura N, Hirakawa H, Sato S, et al. Genome structure of Rosa multiflora, a wild ancestor of cultivated roses [J]. DNA Res, 2018, 25(2): 113-121.
[2]	Zhang Y, Zhao MJ, Zhu W, et al. Nonglandular prickle formation is associated with development and secondary metabolism-related genes in Rosa multiflora [J]. Physiol Plant, 2021, 173(3): 1147-1162.
[3]	McQueen-Mason S, Durachko DM, Cosgrove DJ. Two endogenous proteins that induce cell wall extension in plants [J]. Plant Cell, 1992, 4(11): 1425-1433.
[4]	Arslan B, İncili ÇY, Ulu F, et al. Comparative genomic analysis of expansin superfamily gene members in zucchini and cucumber and their expression profiles under different abiotic stresses [J]. Physiol Mol Biol Plants, 2021, 27(12): 2739-2756.
[5]	Cosgrove DJ. Plant cell wall loosening by expansins [J]. Annu Rev Cell Dev Biol, 2024, 40(1): 329-352.
[6]	Wang ZZ, Cao JB, Lin N, et al. Origin, evolution, and diversification of the expansin family in plants [J]. Int J Mol Sci, 2024, 25(21): 11814.
[7]	Kök BÖ, Celik Altunoglu Y, Öncül AB, et al. Expansin gene family database: a comprehensive bioinformatics resource for plant expansin multigene family [J]. J Bioinform Comput Biol, 2023, 21(3): 2350015.
[8]	Liu WM, Lyu TQ, Xu LA, et al. Complex molecular evolution and expression of expansin gene families in three basic diploid species of Brassica [J]. Int J Mol Sci, 2020, 21(10): 3424.
[9]	Goh HH, Sloan J, Dorca-Fornell C, et al. Inducible repression of multiple expansin genes leads to growth suppression during leaf development [J]. Plant Physiol, 2012, 159(4): 1759-1770.
[10]	Yan A, Wu MJ, Yan LM, et al. AtEXP2 is involved in seed germination and abiotic stress response in Arabidopsis [J]. PLoS One, 2014, 9(1): e85208.
[11]	Feng X, Li CT, He FM, et al. Genome-wide identification of expansin genes in wild soybean (Glycine soja) and functional characterization of Expansin B1 (GsEXPB1) in soybean hair root [J]. Int J Mol Sci, 2022, 23(10): 5407.
[12]	Liu WM, Xu LA, Lin H, et al. Two expansin genes, AtEXPA4 and AtEXPB5, are redundantly required for pollen tube growth and AtEXPA4 is involved in primary root elongation in Arabidopsis thaliana [J]. Genes, 2021, 12(2): 249.
[13]	Jiang FL, Lopez A, Jeon S, et al. Disassembly of the fruit cell wall by the ripening-associated polygalacturonase and expansin influences tomato cracking [J]. Hortic Res, 2019, 6: 17.
[14]	Calderini DF, Castillo FM, Arenas-M A, et al. Overcoming the trade-off between grain weight and number in wheat by the ectopic expression of expansin in developing seeds leads to increased yield potential [J]. New Phytol, 2021, 230(2): 629-640.
[15]	Dai FW, Zhang CQ, Jiang XQ, et al. RhNAC2 and RhEXPA4 are involved in the regulation of dehydration tolerance during the expansion of rose petals [J]. Plant Physiol, 2012, 160(4): 2064-2082.
[16]	Lü PT, Kang M, Jiang XQ, et al. RhEXPA4, a rose expansin gene, modulates leaf growth and confers drought and salt tolerance to Arabidopsis [J]. Planta, 2013, 237(6): 1547-1559.
[17]	Raymond O, Gouzy J, Just J, et al. The Rosagenome provides new insights into the domestication of modern roses [J]. Nat Genet, 2018, 50(6): 772-777.
[18]	Han Y, Yu JY, Zhao T, et al. Dissecting the genome-wide evolution and function of R2R3-MYB transcription factor family in Rosa chinensis [J]. Genes, 2019, 10(10): 823.
[19]	由玉婉, 张雨, 孙嘉毅, 等. ‘月月粉’月季NAC家族全基因组鉴定及皮刺发育相关成员的筛选 [J]. 中国农业科学, 2022, 55(24): 4895-4911.
	You YW, Zhang Y, Sun JY, et al. Genome-wide identification of NAC family and screening of its members related to prickle development in Rosa chinensis old blush [J]. Sci Agric Sin, 2022, 55(24): 4895-4911.
[20]	Chen CJ, Chen H, Zhang Y, et al. TBtools: an integrative toolkit developed for interactive analyses of big biological data [J]. Mol Plant, 2020, 13(8): 1194-1202.
[21]	Hepler NK, Bowman A, Carey RE, et al. Expansin gene loss is a common occurrence during adaptation to an aquatic environment [J]. Plant J, 2020, 101(3): 666-680.
[22]	Krishnamurthy P, Hong JK, Kim JA, et al. Genome-wide analysis of the expansin gene superfamily reveals Brassica rapa-specific evolutionary dynamics upon whole genome triplication [J]. Mol Genet Genomics, 2015, 290(2): 521-530.
[23]	Liu XP, Dong SY, Miao H, et al. Genome-wide analysis of expansins and their role in fruit spine development in cucumber (Cucumis sativus L.) [J]. Hortic Plant J, 2022, 8(6): 757-768.
[24]	Sampedro J, Carey RE, Cosgrove DJ. Genome histories clarify evolution of the expansin superfamily: new insights from the poplar genome and pine ESTs [J]. J Plant Res, 2006, 119(1): 11-21.
[25]	Zhang W, Yan HW, Chen WJ, et al. Genome-wide identification and characterization of maize expansin genes expressed in endosperm [J]. Mol Genet Genomics, 2014, 289(6): 1061-1074.
[26]	Qiu DY, Xu SL, Wang Y, et al. Primary cell wall modifying proteins regulate wall mechanics to steer plant morphogenesis [J]. Front Plant Sci, 2021, 12: 751372.
[27]	Ono K. Signal peptides and their fragments in post-translation: novel insights of signal peptides [J]. Int J Mol Sci, 2024, 25(24): 13534.
[28]	Zhang JL, Dong TT, Zhu MK, et al. Transcriptome- and genome-wide systematic identification of expansin gene family and their expression in tuberous root development and stress responses in sweetpotato (Ipomoea batatas) [J]. Front Plant Sci, 2024, 15: 1412540.
[29]	Lou Y, Zhou HS, Han Y, et al. Positive regulation of AMS by TDF1 and the formation of a TDF1-AMS complex are required for anther development in Arabidopsis thaliana [J]. New Phytol, 2018, 217(1): 378-391.
[30]	Zhou NN, Simonneau F, Thouroude T, et al. Morphological studies of rose prickles provide new insights [J]. Hortic Res, 2021, 8(1): 221.
[31]	Swarnkar MK, Kumar P, Dogra V, et al. Prickle morphogenesis in rose is coupled with secondary metabolite accumulation and governed by canonical MBW transcriptional complex [J]. Plant Direct, 2021, 5(6): e00325.
[32]	Saint-Oyant LH, Ruttink T, Hamama L, et al. A high-quality genome sequence of Rosa chinensis to elucidate ornamental traits [J]. Nat Plants, 2018, 4(7): 473-484.
[33]	Xing SC, Li F, Guo QF, et al. The involvement of an expansin gene TaEXPB23 from wheat in regulating plant cell growth [J]. Biol Plant, 2009, 53(3): 429-434.
[34]	Zou HY, Wenwen YH, Zang GC, et al. OsEXPB2, a β-expansin gene, is involved in rice root system architecture [J]. Mol Breed, 2015, 35(1): 41.
[35]	Lin CF, Choi HS, Cho HT. Root hair-specific EXPANSIN A7 is required for root hair elongation in Arabidopsis [J]. Mol Cells, 2011, 31(4): 393-398.
[36]	Chen SK, Luo YX, Wang GJ, et al. Genome-wide identification of expansin genes in Brachypodium distachyon and functional characterization of BdEXPA27 [J]. Plant Sci, 2020, 296: 110490.