猕猴桃<i>AcSWEET</i>基因家族的鉴定与表达分析

郑帅; 韩世明; 郑浩; 徐明; 苑馨予; 董家琦; 钟彩虹; 张琼

doi:10.11913/PSJ.2095-0837.23329

猕猴桃AcSWEET基因家族的鉴定与表达分析

郑帅^{1, 2,},
韩世明^2,,
郑浩¹,
徐明³,
苑馨予¹,
董家琦^{1, 4},
钟彩虹^1, ,,
张琼^1, ,

1.
中国科学院武汉植物园/中国科学院猕猴桃产业技术工程实验室，武汉 430074
2.
六盘水师范学院生物科学与技术学院，贵州六盘水 553004
3.
陕西省农村科技开发中心，西安 710054
4.
中国科学院大学，北京 100049

基金项目: 中国科学院战略性先导科技专项（XDA24030404）；果树种质创新与利用湖北省重点实验室开放课题（GSSZ202301）；贵州省高等学校猕猴桃资源开发与利用重点实验室项目（黔教技[2022]054）；六盘水市科技计划项目（52020-2020-0906）；陕西省重点研发计划项目（2023-ZDLNY-24）；六盘水师范学院项目（LPSSY2023XKTD09，Lpssyzxxm202304）。

详细信息

作者简介:
郑帅（1996−），男，硕士研究生，研究方向为猕猴桃果树品质（E-mail：1792649816@qq.com）

韩世明（1982−），男，博士，副教授，研究方向为果树育种及栽培技术（E-mail：94684382@qq.com）

通讯作者:
钟彩虹: E-mail：zhongch@wbgcas.cn

张琼: qiongzhang@wbgcas.cn

共同作者

中图分类号: Q943.2
计量
- 文章访问数: 217
- HTML全文浏览量: 50
- PDF下载量: 45
出版历程
- 收稿日期: 2023-11-06
- 录用日期: 2023-12-03
- 刊出日期: 2024-10-30

Identification and expression analysis of the AcSWEET gene family in Actinidia chinensis Planch.

Zheng Shuai^{1, 2,},
Han Shiming^2,,
Zheng Hao¹,
Xu Ming³,
Yuan Xinyu¹,
Dong Jiaqi^{1, 4},
Zhong Caihong^1, ,,
Zhang Qiong^1, ,

1.
Wuhan Botanical Garden, Chinese Academy of Sciences/Chinese Academy of Sciences Engineering Laboratory for Kiwifruit Industrial Technology, Wuhan 430074, China
2.
School of Biological Science and Technology Liupanshui Normal University, Liupanshui, Guizhou 553004, China
3.
Shaanxi Rural Science and Technology Development Center, Xi’an 710054, China
4.
University of Chinese Academy of Sciences, Beijing 100049, China

摘要

摘要:
SWEET（Sugars will eventually be exported transporters）是近年来在植物中发现的一组糖转运蛋白，在植物生长、发育和非生物及生物胁迫响应等多种生理过程中发挥着重要作用。本研究利用生物信息学方法对猕猴桃（Actinidia chinensis Planch.）AcSWEET基因家族进行了鉴定，共获得29个AcSWEET基因，并对其氨基酸数量、相对分子量、等电点、不稳定系数、亚细胞定位、亲水指数进行了分析。结果显示：29个基因编码的氨基酸数目为680~906个；分子量范围为7.531~101.266 kDa，等电点在6.95~9.90，多数蛋白为定位于细胞膜的疏水性蛋白，具有1~2个MtN3结构域或PQ-loop结构域。此外，AcSWEET基因的外显子数量在4~6个，系统进化分析结果表明猕猴桃AcSWEET基因家族被分为4个亚族，同一亚族基因具有相似的内含子、外显子以及保守基序。表达模式分析结果表明，这些基因在果实不同发育时期具有表达特异性。推测AcSWEET26、AcSWEET7、AcSWEET15和AcSWEET13可能参与猕猴桃的蔗糖转运和积累。
- 猕猴桃 /
- SWEET基因 /
- 基因家族 /
- 基因表达分析
Abstract:
The sugar will eventually be exported transporters (SWEET) are a recently discovered group of sugar transporters in plants, which play important roles in various physiological processes, such as plant growth, development, and abiotic and biotic stress. This study aimed to characterize the Actinidia chinensis Planch. SWEET gene family and analyze its expression during fruit development using bioinformatics methods. A total of 29 AcSWEET genes were identified, and their amino acid quantity, relative molecular weights, isoelectric points, instability coefficients, subcellular localizations, and hydrophilicity indices were analyzed. Results showed that the 29 AcSWEET genes encoded proteins with amino acid lengths ranging from 680 to 906 residues, molecular weights between 7.531 kDa and 101.266 kDa, and isoelectric points between 6.95 and 9.9. Most proteins were relatively stable hydrophobic, and localized on the cell membrane, containing 1–2 MtN3 domains. Additionally, gene structure, conserved motifs, evolutionary relationships, cis-regulatory elements, and expression patterns at different developmental stages of fruit development were systematically analyzed. Results indicated that the AcSWEET genes had 4–6 exons. Phylogenetic analysis revealed that the kiwifruit SWEET gene family was divided into four subgroups, responsible for transporting glucose, fructose, and sucrose. Genes within the same subgroup shared similar intron-exon structures and conserved motifs. Expression pattern analysis during kiwifruit development indicated that the AcSWEET genes exhibited stage-specific expression. We speculated that AcSWEET26, AcSWEET7, AcSWEET15, and AcSWEET13 may be involved in sucrose transport and accumulation in kiwifruit.
- Actinidia chinensis /
- SWEET gene /
- Gene family /
- Gene expression analysis
21~2）如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

3如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

1如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

4如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

5如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

HTML全文

21~2）如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

3如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

1如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

4如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

5如需查阅附图内容请登录《植物科学学报》网站（http://www.plantscience.cn）查看本期文章。

图 1 AcSWEET基因在染色体上的分布

Figure 1. Distribution of AcSWEET genes on chromosomes

下载: 全尺寸图片幻灯片

图 2 SWEET基因家族系统进化树

Figure 2. Phylogenetic tree of SWEET gene family

下载: 全尺寸图片幻灯片

图 3 AcSWEET基因家族启动子顺式作用元件

Figure 3. Cis-regulatory elements of AcSWEET gene family promoters

下载: 全尺寸图片幻灯片

图 4 不同发育阶段猕猴桃AcSWEET基因家族表达量分析

Figure 4. Expression levels of AcSWEET gene family at different developmental stages

下载: 全尺寸图片幻灯片

图 5 AcSWEET基因在猕猴桃果实不同发育时间的表达分析

A为FPKM值， B为荧光定量PCR结果。不同小写字母表示在P<0.05水平上差异显著。

Figure 5. Expression analysis of AcSWEET genes at different developmental stages

A shows FPKM values, B shows result of qRT-PCR. Different lowercase letters indicate significant differences at P<0.05 level.

下载: 全尺寸图片幻灯片

图 6 ‘东红’猕猴桃果实蔗糖、果糖及葡萄糖含量

Figure 6. Sucrose, fructose and glucose contents in ‘Donghong’ fruit

下载: 全尺寸图片幻灯片

表 1 实时荧光定量PCR引物

Table 1 Quantitative real-time polymerase chain reaction primers

基因名称 Gene name	正向引物（5′–3′） Forward primer	反向引物（5′–3′） Reverse primer
AcSWEET7	TAGTCGTTGGATGGGTTTGC	TGCGGTCATTGTAAGGAAAA
AcSWEET13	ATAAGAAGAAATCAACCGAAGG	GCAGCAAATACGCTCACAGA
AcSWEET15	GAGTCACGATTGTCGGATGG	TTGGAGCTTCACTTCTGGTAGA
AcSWEET26	CACCCTTCTCATCACCATCA	TAGGCAAATCCAACCAACAA
AcSWEET29	CACATCCCTATTATCAAACG	AAATTCTGGTAGAACCTGCT
Actin	GTGCTCAGTGGTGGTTCAA	GACGCTGTATTTCCTCTCAG

下载: 导出CSV

表 2 AcSWEET基因家族成员信息

Table 2 Information on AcSWEET gene family members

基因 Gene	序列ID Sequence ID	蛋白质长度 Number of amino acids	分子量 MW / Da	等电点 Theoretical pI	不稳定系数 Instability index	脂溶指数 Aliphatic index	亲水指数 Hydrophobicity scales	亚细胞定位 Subcellular localization	染色体位置 Chromosome location
AcSWEET1	Ach05g06950DH	68	7 531.83	8.03	29.09	117.65	0.469	叶绿体	5
AcSWEET2	Ach06g07770DH	245	2 7196.90	6.95	47.57	103.39	0.568	质膜	6
AcSWEET3	Ach06g08520DH	236	26 023.09	9.36	35.65	119.36	0.876	液泡膜	6
AcSWEET4	Ach10g04100DH	253	27 658.05	9.57	29.51	114.82	0.590	质膜	10
AcSWEET5	Ach10g10290DH	236	26 340.97	9.31	35.04	95.42	0.163	细胞核	10
AcSWEET6	Ach11g04370DH	252	27 746.95	9.37	34.26	109.44	0.512	质膜	11
AcSWEET7	Ach13g04100DH	288	32 042.07	6.99	36.66	120.14	0.694	质膜	13
AcSWEET8	Ach13g17010DH	234	25 814.53	8.80	36.02	111.58	0.684	质膜	13
AcSWEET9	Ach14g03170DH	198	21 765.16	9.71	29.99	120.51	0.703	质膜	14
AcSWEET10	Ach14g03210DH	186	20 525.26	9.91	31.15	117.96	0.334	质膜	14
AcSWEET11	Ach16g04520DH	103	11 850.58	4.93	40.50	73.79	−0.449	细胞质	16
AcSWEET12	Ach16g04530DH	136	15 558.83	9.44	31.10	129.78	0.856	质膜	16
AcSWEET13	Ach16g04540DH	287	32 139.66	9.39	38.65	117.46	0.778	质膜	16
AcSWEET14	Ach16g06160DH	238	26 764.96	9.10	37.01	115.46	0.671	质膜	16
AcSWEET15	Ach17g10410DH	275	30 968.82	7.61	35.74	111.35	0.656	质膜	17
AcSWEET16	Ach17g10420DH	132	14 617.52	9.08	49.24	108.71	0.679	叶绿体	17
AcSWEET17	Ach18g07250DH	250	28 204.71	9.37	44.39	115.76	0.554	质膜	18
AcSWEET18	Ach19g02550DH	296	33 224.04	8.46	35.78	130.57	0.841	质膜	19
AcSWEET19	Ach19g10330DH	304	33 198.18	9.49	29.29	111.48	0.346	质膜	19
AcSWEET20	Ach19g11300DH	906	101 266.08	9.61	46.80	86.35	−0.272	质膜	19
AcSWEET21	Ach21g07220DH	252	27 533.77	9.59	28.91	114.17	0.663	质膜	21
AcSWEET22	Ach23g12130DH	235	26 009.82	9.30	45.47	118.64	0.802	液泡膜	23
AcSWEET23	Ach24g12310DH	244	27 010.81	8.48	35.30	116.23	0.618	质膜	24
AcSWEET24	Ach25g05410DH	259	28 487.96	9.63	43.00	115.52	0.558	液泡膜	25
AcSWEET25	Ach26g07610DH	183	20 725.66	8.49	41.84	114.48	0.680	液泡膜	26
AcSWEET26	Ach26g09140DH	290	32 463.83	8.77	38.39	114.93	0.722	质膜	26
AcSWEET27	Ach26g09150DH	340	38 910.41	9.24	48.76	88.85	0.107	质膜	26
AcSWEET28	Ach27g01830DH	81	9 064.60	9.30	39.94	101.11	0.251	叶绿体	27
AcSWEET29	Ach28g11120DH	261	29 612.67	8.97	37.55	117.36	0.777	质膜	28
注：亲水指数为负值表示亲水性，正值表示疏水性；不稳定指数大于 40 为不稳定蛋白；脂溶指数小于 100 为脂溶蛋白。
Notes: For total average hydrophilicity, a negative value indicates hydrophilicity, while a positive value indicates hydrophobicity; Instability index greater than 40 indicates an unstable protein; Lipid solubility index less than 100 indicates lipid soluble protein.

下载: 导出CSV

表 3 猕猴桃果实发育过程中糖含量及候选基因的相关性

Table 3 Correlation of sugar content and candidate genes at different developmental stages

指标 Item	蔗糖 Sucrose	葡萄糖 Glucose	果糖 Fructose	AcSWEET7	AcSWEET13	AcSWEET15	AcSWEET26	AcSWEET29
蔗糖	1
葡萄糖	0.960**	1
果糖	0.977**	0.975**	1
AcSWEET7	0.754**	0.664*	0.751**	1
AcSWEET13	0.677*	0.698*	0.694*	0.466	1
AcSWEET15	0.773**	0.686*	0.776**	0.972**	0.411	1
AcSWEET26	0.541	0.467	0.549	0.542	0.397	0.693*	1
AcSWEET29	0.904**	0.836**	0.915**	0.910**	0.566	0.934**	0.647*	1
Notes: : P<0.05; *: P<0.01.

下载: 导出CSV

参考文献(35)

[1]	黄宏文. 中国猕猴桃种质资源[M]. 北京:中国林业出版社,2013:10−27.
[2]	Zhang L,Tang ZM,Zheng H,Zhong CH,Zhang Q. Comprehensive analysis of metabolome and transcriptome in fruits and roots of kiwifruit[J]. Int J Mol Sci,2023,24(2):1299. doi: 10.3390/ijms24021299
[3]	Ruan YL. Sucrose metabolism:gateway to diverse carbon use and sugar signaling[J]. Annu Rev Plant Biol,2014,65:33−67. doi: 10.1146/annurev-arplant-050213-040251
[4]	Schulz A,Beyhl D,Marten I,Wormit A,Neuhaus E,et al. Proton-driven sucrose symport and antiport are provided by the vacuolar transporters SUC4 and TMT1/2[J]. Plant J,2011,68(1):129−136. doi: 10.1111/j.1365-313X.2011.04672.x
[5]	Kühn C,Grof CPL. Sucrose transporters of higher plants[J]. Curr Opin Plant Biol,2010,13(3):287−297. doi: 10.1016/j.pbi.2010.02.001
[6]	Ayre BG. Membrane-transport systems for sucrose in relation to whole-plant carbon partitioning[J]. Mol Plant,2011,4(3):377−394. doi: 10.1093/mp/ssr014
[7]	Chen LQ,Hou BH,Lalonde S,Takanaga H,Hartung ML,et al. Sugar transporters for intercellular exchange and nutrition of pathogens[J]. Nature,2010,468(7323):527−532. doi: 10.1038/nature09606
[8]	Doidy J,Grace E,Kühn C,Simon-Plas F,Casieri L,Wipf D. Sugar transporters in plants and in their interactions with fungi[J]. Trends Plant Sci,2012,17(7):413−422. doi: 10.1016/j.tplants.2012.03.009
[9]	Chen LQ. SWEET sugar transporters for phloem transport and pathogen nutrition[J]. New Phytol,2014,201(4):1150−1155. doi: 10.1111/nph.12445
[10]	Eom JS,Chen LQ,Sosso D,Julius BT,Lin IW,et al. SWEETs,transporters for intracellular and intercellular sugar translocation[J]. Curr Opin Plant Biol,2015,25:53−62. doi: 10.1016/j.pbi.2015.04.005
[11]	Hamada M,Wada S,Kobayashi K,Satoh N. Ci-Rga,a gene encoding an MtN3/saliva family transmembrane protein,is essential for tissue differentiation during embryogenesis of the ascidian Ciona intestinalis[J]. Differentiation,2005,73(7):364−376. doi: 10.1111/j.1432-0436.2005.00037.x
[12]	Xuan YH,Hu YB,Chen LQ,Sosso D,Ducat DC,et al. Functional role of oligomerization for bacterial and plant SWEET sugar transporter family[J]. Proc Natl Acad Sci USA,2013,110(39):E3685−E3694.
[13]	Liu HT,Lyu WY,Tian SH,Zou XH,Zhang LQ,et al. The SWEET family genes in strawberry:identification and expression profiling during fruit development[J]. South Afr J Bot,2019,125:176−187. doi: 10.1016/j.sajb.2019.07.002
[14]	Chen LQ,Qu XQ,Hou BH,Sosso D,Osorio S,et al. Sucrose efflux mediated by SWEET proteins as a key step for phloem transport[J]. Science,2012,335(6065):207−211. doi: 10.1126/science.1213351
[15]	Chardon F,Bedu M,Calenge F,Klemens PAW,Spinner L,et al. Leaf fructose content is controlled by the vacuolar transporter SWEET17 in Arabidopsis[J]. Curr Biol,2013,23(8):697−702. doi: 10.1016/j.cub.2013.03.021
[16]	Gao Y,Wang ZY,Kumar V,Xu XF,Yuan DP,et al. Genome-wide identification of the SWEET gene family in wheat[J]. Gene,2018,642:284−292. doi: 10.1016/j.gene.2017.11.044
[17]	Feng CY,Han JX,Han XX,Jiang J. Genome-wide identification,phylogeny,and expression analysis of the SWEET gene family in tomato[J]. Gene,2015,573(2):261−272. doi: 10.1016/j.gene.2015.07.055
[18]	Li W,Ren ZY,Wang ZY,Sun K,Pei XY,et al. Evolution and stress responses of Gossypium hirsutum SWEET genes[J]. Int J Mol Sci,2018,19(3):769. doi: 10.3390/ijms19030769
[19]	Guo CY,Li HY,Xia XY,Liu XY,Yang L. Functional and evolution characterization of SWEET sugar transporters in Ananas comosus[J]. Biochem Biophys Res Commun,2018,496(2):407−414. doi: 10.1016/j.bbrc.2018.01.024
[20]	Zhu JL,Zhou L,Li TF,Ruan YY,Zhang A,et al. Genome-wide investigation and characterization of SWEET gene family with focus on their evolution and expression during hormone and abiotic stress response in maize[J]. Genes,2022,13(10):1682. doi: 10.3390/genes13101682
[21]	Dai ZR,Yan PY,He SZ,Jia LC,Wang YN,et al. Genome-wide identification and expression analysis of SWEET family genes in sweet potato and its two diploid relatives[J]. Int J Mol Sci,2022,23(24):15848. doi: 10.3390/ijms232415848
[22]	Yao TS,Xie RJ,Zhou Y,Hu JH,Gao Y,Zhou CY. Genome-wide identification of SWEET gene family and its response to abiotic stresses in Valencia sweet orange[J]. Plant Mol Biol Rep,2021,39(3):546−556. doi: 10.1007/s11105-020-01268-1
[23]	Jiang SJ,Balan B,de A. B. Assis R,Sagawa CHD,Wan XQ,et al. Genome-wide profiling and phylogenetic analysis of the SWEET sugar transporter gene family in walnut and their lack of responsiveness to Xanthomonas arboricola pv. juglandis infection[J]. Int J Mol Sci,2020,21(4):1251. doi: 10.3390/ijms21041251
[24]	Hu LP,Zhang F,Song SH,Tang XW,Xu H,et al. Genome-wide identification,characterization,and expression analysis of the SWEET gene family in cucumber[J]. J Integr Agric,2017,16(7):1486−1501. doi: 10.1016/S2095-3119(16)61501-0
[25]	Li JM,Qin MF,Qiao X,Cheng YS,Li XL,et al. A new insight into the evolution and functional divergence of sweet transporters in Chinese white pear (Pyrus bretschneideri)[J]. Plant Cell Physiol,2017,58(4):839−850. doi: 10.1093/pcp/pcx025
[26]	Wen ZY,Li MY,Meng J,Li P,Cheng TR,et al. Genome-wide identification of the SWEET gene family mediating the cold stress response in Prunus mume[J]. PeerJ,2022,10:e13273. doi: 10.7717/peerj.13273
[27]	Sui JL,Xiao XH,Qi JY,Fang YJ,Tang CR. The SWEET gene family in Hevea brasiliensis-its evolution and expression compared with four other plant species[J]. FEBS Open Bio,2017,7(12):1943−1959. doi: 10.1002/2211-5463.12332
[28]	Zhang XH,Wang S,Ren Y,Gan CY,Li BB,et al. Identification,analysis and gene cloning of the SWEET gene family provide insights into sugar transport in pomegranate (Punica granatum)[J]. Int J Mol Sci,2022,23(5):2471. doi: 10.3390/ijms23052471
[29]	Fang T,Rao Y,Wang MZ,Li Y,Liu YJ,et al. Characterization of the SWEET gene family in longan (Dimocarpus longan) and the role of DlSWEET1 in cold tolerance[J]. Int J Mol Sci,2022,23(16):8914. doi: 10.3390/ijms23168914
[30]	Han X,Zhang YL,Zhang Q,Ma N,Liu XY,et al. Two haplotype-resolved,gap-free genome assemblies for Actinidia latifolia and Actinidia chinensis shed light on the regulatory mechanisms of vitamin C and sucrose metabolism in kiwifruit[J]. Mol Plant,2023,16(2):452−470. doi: 10.1016/j.molp.2022.12.022
[31]	Chen CJ,Chen H,Zhang Y,Thomas HR,Frank MH,et al. TBtools:an integrative toolkit developed for interactive analyses of big biological data[J]. Mol Plant,2020,13(8):1194−1202. doi: 10.1016/j.molp.2020.06.009
[32]	Ampomah-Dwamena C,McGhie T,Wibisono R,Montefiori M,Hellens RP,Allan AC. The kiwifruit lycopene beta-cyclase plays a significant role in carotenoid accumulation in fruit[J]. J Exp Bot,2009,60(13):3765−3779. doi: 10.1093/jxb/erp218
[33]	Zhao K,Chen S,Yao WJ,Cheng ZH,Zhou BR,Jiang TB. Genome-wide analysis and expression profile of the bZIP gene family in poplar[J]. BMC Plant Biol,2021,21(1):122. doi: 10.1186/s12870-021-02879-w
[34]	Yang JJ,Zhan RL,Jin YR,Song JY,Li DX,et al. Functional analysis of the promoter of the MdFRK2 gene encoding a high-affinity fructokinase in apple (Malus × domestica)[J]. Sci Hortic,2020,265:109088. doi: 10.1016/j.scienta.2019.109088
[35]	Xu XH,Li WL,Yang SK,Zhu XZ,Sun HW,et al. Identification,evolution,expression and protein interaction analysis of genes encoding B-box zinc-finger proteins in maize[J]. J Integr Agric,2023,22(2):371−388. doi: 10.1016/j.jia.2022.08.091