猕猴桃AcHSP20基因家族的鉴定及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2023-1092

摘要/Abstract

摘要：

【目的】鉴定分析了猕猴桃的小热休克蛋白（HSP20s/sHSPs）基因家族，为猕猴桃AcHSP20基因的生物学功能研究和猕猴桃的抗性育种奠定基础。【方法】基于猕猴桃全基因组数据，利用生物信息学方法对猕猴桃AcHSP20基因家族进行鉴定，并分析了家族成员的理化性质、系统进化、染色体定位、基因结构、亚细胞定位及启动子。同时利用荧光定量PCR技术分析了猕猴桃AcHSP20基因在非生物胁迫和激素处理后的表达变化情况。【结果】鉴定获得34个AcHSP20基因家族成员。其编码蛋白的氨基酸数目为85-371，分子量介于9.93-40.18 kD，等电点介于4.4-9.3，AcHSP20s多定位于细胞质和叶绿体。34个基因分布在17条染色体上，多数没有或只有一个内含子。基因的启动子含有植物激素和非生物胁迫响应元件。所有测定的AcHSP20基因在猕猴桃的根茎叶中均有表达且在高温及其他多种非生物胁迫和植物激素处理下差异表达显著。【结论】AcHSP20基因家族成员在猕猴桃的高温、ABA、MeJA、NaCl等逆境胁迫响应中发挥重要作用。

关键词: 猕猴桃, AcHSP20基因家族, 基因特征, 生物信息学

Abstract:

【Objective】The identification and analyses of heat shock proteins（HSP20s/sHSPs）in kiwifruit would lay foundation for the functional studies of AcHSP20 genes and resistance breeding of kiwifruit. 【Method】Based on the genome-wide data of kiwifruit, the AcHSP20 gene family was identified, and the physicochemical properties, phyletic evolution, chromosome distribution, gene structure, subcellular localization as well as promoters were analyzed by bioinformatics. 【Result】The 34 members of AcHSP20 gene family were identified. They encoded proteins with 85-371 amino acids, 9.93-40.18 kD molecular weight（Mw）and 4.4-9.3 isoelectric points（pI）. Meanwhile, most AcHSP20 proteins were predicted to be located in cytoplasm and chloroplast. The 34 AcHSP20 genes were distributed on 17 chromosomes, and there were no or only one intron, mostly. Promoters of AcHSP20 genes had phytohormone and abiotic stress response elements. All the detected AcHSP20 genes expressed ubiquitously in the roots, stems and leaves of kiwifruit and the expressions changed significantly under abiotic stresses and phytohormone treatments. 【Conclusion】The AcHSP20 gene family plays important roles in high temperature, ABA, MeJA, NaCl stress responses in kiwifruit.

Key words: kiwifruit, AcHSP20 gene family, gene characterization, bioinformatics

侯雅琼, 郎红珊, 闻蒙蒙, 谷易云, 朱润洁, 汤晓丽. 猕猴桃AcHSP20基因家族的鉴定及表达分析[J]. 生物技术通报, 2024, 40(5): 167-178.

HOU Ya-qiong, LANG Hong-shan, WEN Meng-meng, GU Yi-yun, ZHU Run-jie, TANG Xiao-li. Identification and Expression Analysis of AcHSP20 Gene Family in Kiwifruit[J]. Biotechnology Bulletin, 2024, 40(5): 167-178.

图/表 9

参考文献 40

[1]	Herman DJ, Knowles LO, Knowles NR. Heat stress affects carbohydrate metabolism during cold-induced sweetening of potato(Solanum tuberosum L.)[J]. Planta, 2017, 245(3): 563-582. doi: 10.1007/s00425-016-2626-z pmid: 27904974
[2]	Giorno F, Wolters-Arts M, Grillo S, et al. Developmental and heat stress-regulated expression of HsfA2 and small heat shock proteins in tomato anthers[J]. J Exp Bot, 2010, 61(2): 453-462. doi: 10.1093/jxb/erp316 pmid: 19854799
[3]	Peleg Z, Blumwald E. Hormone balance and abiotic stress tolerance in crop plants[J]. Curr Opin Plant Biol, 2011, 14(3): 290-295. doi: 10.1016/j.pbi.2011.02.001 pmid: 21377404
[4]	Wang N, Guo TL, Sun X, et al. Functions of two Malus hupehensis(Pamp.) Rehd. YTPs(MhYTP1 and MhYTP2)in biotic- and abiotic-stress responses[J]. Plant Sci, 2017, 261: 18-27. doi: S0168-9452(17)30238-8 pmid: 28554690
[5]	Suzuki N. Temperature stress and responses in plants[J]. Int J Mol Sci, 2019, 20(8): 2001.
[6]	Yang GD, Yu ZP, Gao L, et al. SnRK2s at the crossroads of growth and stress responses[J]. Trends Plant Sci, 2019, 24(8): 672-676. doi: S1360-1385(19)30139-6 pmid: 31255544
[7]	Tang XL, Mu XM, Shao HB, et al. Global plant-responding mechanisms to salt stress: physiological and molecular levels and implications in biotechnology[J]. Crit Rev Biotechnol, 2015, 35(4): 425-437. doi: 10.3109/07388551.2014.889080 pmid: 24738851
[8]	Tang XL, Shao HB, Jiang FD, et al. Molecular cloning and functional analyses of the salt-responsive gene KvHSP70 from Kosteletzkya virginica[J]. Land Degrad Dev, 2020, 31(6): 773-782.
[9]	Wu JT, Gao T, Hu JN, et al. Research advances in function and regulation mechanisms of plant small heat shock proteins(sHSPs)under environmental stresses[J]. Sci Total Environ, 2022, 825: 154054.
[10]	Jacob P, Hirt H, Bendahmane A. The heat-shock protein/chaperone network and multiple stress resistance[J]. Plant Biotechnol J, 2017, 15(4): 405-414. doi: 10.1111/pbi.12659 pmid: 27860233
[11]	Zhang KM, Ezemaduka AN, Wang Z, et al. A novel mechanism for small heat shock proteins to function as molecular chaperones[J]. Sci Rep, 2015, 5(5): 8811-8819.
[12]	Haq S U, Khan A, Ali M, et al. Heat shock proteins: Dynamic biomolecules to counter plant biotic and abiotic stresses[J]. Int J Mol Sci, 2019, 20(21): 5321.
[13]	Tian C, Zhang ZY, Huang Y, et al. Functional characterization of the Pinellia ternata cytoplasmic class II small heat shock protein gene PtsHSP17.2 via promoter analysis and overexpression in tobacco[J]. Plant Physiol Biochem, 2022, 177: 1-9.
[14]	Feng XH, Zhang HX, Ali M, et al. A small heat shock protein CaHsp25.9 positively regulates heat, salt, and drought stress tolerance in pepper(Capsicum annuum L.)[J]. Plant Physiol Biochem, 2019, 142(142): 151-162.
[15]	Waters ER. The evolution, function, structure and expression of the plant sHSPs[J]. J Exp Bot, 2013, 64(2): 391-403.
[16]	Waters ER, Vierling E. Plant small heat shock proteins-evolutionary and functional diversity[J]. New Phytol, 2020, 227(1): 24-37.
[17]	Haslbeck M, Franzmann T, Weinfurtner D, et al. Some like it hot: the structure and function of small heat-shock proteins[J]. Nat Struct Mol Biol, 2005, 12(10): 842-846. doi: 10.1038/nsmb993 pmid: 16205709
[18]	El-Gebali S, Mistry J, Bateman A, et al. The Pfam protein families database in 2019[J]. Nucleic Acids Res, 2019, 47(D1): D427-D432.
[19]	Ma W, Guan XY, Li J, et al. Mitochondrial small heat shock protein mediates seed germination via thermal sensing[J]. Proc Natl Acad Sci USA, 2019, 116(10): 4716-4721. doi: 10.1073/pnas.1815790116 pmid: 30765516
[20]	Rutsdottir G, Härmark J, Weide Y, et al. Structural model of dodecameric heat-shock protein Hsp21: Flexible N-terminal arms interact with client proteins while C-terminal tails maintain the dodecamer and chaperone activity[J]. J Biol Chem, 2017, 292(19): 8103-8121. doi: 10.1074/jbc.M116.766816 pmid: 28325834
[21]	Liberek K, Lewandowska A, Zietkiewicz S. Chaperones in control of protein disaggregation[J]. EMBO J, 2008, 27(2): 328-335. doi: 10.1038/sj.emboj.7601970 pmid: 18216875
[22]	Yang ML, Zhang YX, Zhang HH, et al. Identification of MsHsp20 gene family in malus sieversii and functional characterization of MsHsp16.9 in heat tolerance[J]. Front Plant Sci, 2017, 8: 1761-1778.
[23]	Sewelam N, Kazan K, Hüdig M, et al. The AtHSP17.4C1 gene expression is mediated by diverse signals that link biotic and abiotic stress factors with ROS and can be a useful molecular marker for oxidative stress[J]. Int J Mol Sci, 2019, 20(13): 3201-3218.
[24]	Mu CJ, Zhang SJ, Yu GZ, et al. Overexpression of small heat shock protein LimHSP16.45 in Arabidopsis enhances tolerance to abiotic stresses[J]. PLoS One, 2013, 8(12): e82264-e82273.
[25]	Zhang L, Gao YK, Pan HT, et al. Cloning and characterisation of a Primula heat shock protein gene, PfHSP17.1, which confers heat, salt and drought tolerance in transgenic Arabidopsis thaliana[J]. Acta Physiol Plant, 2013, 35(11): 3191-3200.
[26]	Sun XB, Sun CY, Li ZG, et al. AsHSP17, a creeping bentgrass small heat shock protein modulates plant photosynthesis and ABA-dependent and independent signalling to attenuate plant response to abiotic stress[J]. Plant Cell Environ, 2016, 39(6): 1320-1337.
[27]	Zhang N, Zhao HY, Shi JW, et al. Functional characterization of class I SlHSP17.7 gene responsible for tomato cold-stress tolerance[J]. Plant Sci, 2020, 298: 110568.
[28]	Do VG, Lee Y, Kweon H, et al. Light induces carotenoid biosynthesis-related gene expression, accumulation of pigment content, and expression of the small heat shock protein in apple fruit[J]. Int J Mol Sci, 2022, 23(11): 6153.
[29]	Kaur H, Petla BP, Kamble NU, et al. Differentially expressed seed aging responsive heat shock protein OsHSP18.2 implicates in seed vigor, longevity and improves germination and seedling establishment under abiotic stress[J]. Front Plant Sci, 2015, 6: 713. doi: 10.3389/fpls.2015.00713 pmid: 26442027
[30]	Kuang J, Liu JZ, Mei J, et al. A class II small heat shock protein OsHsp18.0 plays positive roles in both biotic and abiotic defense responses in rice[J]. Sci Rep, 2017, 7(1): 11333. doi: 10.1038/s41598-017-11882-x pmid: 28900229
[31]	Abassi S, Wang H, Ponmani T, et al. Small heat shock protein genes of the green algae Closterium ehrenbergii: Cloning and differential expression under heat and heavy metal stresses[J]. Environ Toxicol, 2019, 34(9): 1013-1024. doi: 10.1002/tox.22772 pmid: 31095847
[32]	Li ZY, Long RC, Zhang TJ, et al. Molecular cloning and characterization of the MsHSP17.7 gene from Medicago sativa L.[J]. Mol Biol Rep, 2016, 43(8): 815-826.
[33]	Li XW, Li JQ, Djaja DS. New synonyms in Actinidiaceae from China[J]. J Syst Evol, 2007, 45(5): 633-660.
[34]	钟彩虹, 黄文俊, 李大卫, 等. 世界猕猴桃产业发展及鲜果贸易动态分析[J]. 中国果树, 2021, 7(7): 101-108.
	Zhong CH, Huang WJ, Li DW, et al. Dynamic analysis of global kiwifruit industry development and fresh fruit trade[J]. China Fruits, 2021, 7(7): 101-108.
[35]	Hua YG, Liu Q, Zhai YF, et al. Genome-wide analysis of the HSP20 gene family and its response to heat and drought stress in Coix(Coix lacryma-jobi L.)[J]. BMC Genomics, 2023, 24(1): 478-494.
[36]	Scharf KD, Siddique M, Vierling E. The expanding family of Arabidopsis thaliana small heat stress proteins and a new family of proteins containing alpha-crystallin domains(Acd proteins)[J]. Cell Stress Chaperones, 2001, 6(3): 225-237.
[37]	Ouyang YD, Chen JJ, Xie WB, et al. Comprehensive sequence and expression profile analysis of Hsp20 gene family in rice[J]. Plant Mol Biol, 2009, 70(3): 341-357. doi: 10.1007/s11103-009-9477-y pmid: 19277876
[38]	Mattick JS, Gagen MJ. The evolution of controlled multitasked gene networks: the role of introns and other noncoding RNAs in the development of complex organisms[J]. Mol Biol Evol, 2001, 18(9): 1611-1630. doi: 10.1093/oxfordjournals.molbev.a003951 pmid: 11504843
[39]	Huang LJ, Cheng GX, Khan A, et al. CaHSP16.4, a small heat shock protein gene in pepper, is involved in heat and drought tolerance[J]. Protoplasma, 2019, 256(1): 39-51.
[40]	He YJ, Yao YX, Li LL, et al. A heat-shock 20 protein isolated from watermelon(ClHSP22.8)negatively regulates the response of Arabidopsis to salt stress via multiple signaling pathways[J]. PeerJ, 2021, 9: e10524.

基因Gene	上游引物Forward primer（5'-3'）	下游引物Reverse primer（5'-3'）
AcHSP20-2	GATCTATCGCACCGTGTC	AACCTTTATGTCGTCCTTG
AcHSP20-5	CTTCCAGGAGGTGTATCGT	CGTCTTCTTTGGCATTGTA
AcHSP20-9	TCTCACCTTTGGGTTTATTG	CAGGGAACGTCATGGAA
AcHSP20-12	CGGGAGGAGGAGAAAGAA	CAAACAGCAGACACGGC
AcHSP20-19	AGAAGAATGGAGGTGATGAT	GAGATGGAGCACAAGAGTTT
AcActin	TGCATGAGCGATCAAGTTTCAAG	TGTCCCATGTCTGGTTGATGACT

基因Gene	上游引物Forward primer（5'-3'）	下游引物Reverse primer（5'-3'）
AcHSP20-2	GATCTATCGCACCGTGTC	AACCTTTATGTCGTCCTTG
AcHSP20-5	CTTCCAGGAGGTGTATCGT	CGTCTTCTTTGGCATTGTA
AcHSP20-9	TCTCACCTTTGGGTTTATTG	CAGGGAACGTCATGGAA
AcHSP20-12	CGGGAGGAGGAGAAAGAA	CAAACAGCAGACACGGC
AcHSP20-19	AGAAGAATGGAGGTGATGAT	GAGATGGAGCACAAGAGTTT
AcActin	TGCATGAGCGATCAAGTTTCAAG	TGTCCCATGTCTGGTTGATGACT

基因 Gene	基因组登录号 Gene ID	等电点 pI	分子量 Molecular weight/kD	氨基酸数Number of amino acids	染色体定位及基因方向Chromosome localization and gene orientation	亚细胞定位 Subcellular location
AcHSP20-1	Ach00g218421.2	6.57	10.16	88	Chr00: 54153215.. 54154904（-）	细胞质Cytoplasmic
AcHSP20-2	Ach00g239181	7.86	16.14	143	Chr00: 50739806.. 50740562（-）	过氧化物酶体Peroxisomal
AcHSP20-3	Ach00g329621	5.59	23.28	208	Chr00: 90841963.. 90843153（-）	叶绿体Chloroplast
AcHSP20-4	Ach00g479741.2	10.32	9.93	86	Chr00: 135211617.. 135211877（+）	细胞质Cytoplasmic
AcHSP20-5	Ach00g479761.2	9.14	40.18	371	Chr00: 145805825.. 145808080（-）	高尔基体Golgi apparatus
AcHSP20-6	Ach02g036831	7.8	16.56	152	Chr02: 4423490.. 4423948（-）	细胞质Cytoplasmic
AcHSP20-7	Ach02g046741.2	6.21	25.55	225	Chr02: 516872.. 518080（-）	叶绿体Chloroplast
AcHSP20-8	Ach02g223291	8.86	17.61	153	Chr02: 4090770.. 4091231（-）	细胞质Cytoplasmic
AcHSP20-9	Ach03g045661	7.63	25.69	232	Chr03: 6797048.. 6798489（-）	叶绿体Chloroplast
AcHSP20-10	Ach03g451751.2	9.15	11.12	96	Chr03: 1045510.. 1045800（-）	叶绿体Chloroplast
AcHSP20-11	Ach04g161211.2	6.45	24.77	215	Chr04: 6179073.. 6180486（-）	细胞质Cytoplasmic
AcHSP20-12	Ach06g418911.2	6.32	16.60	152	Chr06: 2925504.. 2925962（-）	细胞质Cytoplasmic
AcHSP20-13	Ach10g027441.2	5.4	20.33	181	Chr10: 4989172.. 4989717（-）	细胞质Cytoplasmic
AcHSP20-14	Ach10g027451	7.77	25.72	228	Chr10: 4973842.. 4974528（-）	细胞质Cytoplasmic
AcHSP20-15	Ach10g027461.2	6.34	24.95	221	Chr10: 4984511.. 4985176（+）	叶绿体Chloroplast
AcHSP20-16	Ach10g143161	9.3	25.78	228	Chr10: 11814664.. 11815350（+）	细胞质Cytoplasmic
AcHSP20-17	Ach11g263431.2	5.05	21.72	196	Chr11: 8776984.. 8777574（-）	细胞质Cytoplasmic
AcHSP20-18	Ach11g425231.2	4.43	10.59	90	Chr11: 12263315.. 12263587（-）	细胞质Cytoplasmic
AcHSP20-19	Ach12g058351.2	8.69	17.21	152	Chr12: 6955029.. 6956940（+）	细胞核Nucleus
AcHSP20-20	Ach13g308811	5.94	33.07	297	Chr13: 1116598.. 1117580（+）	叶绿体Chloroplast
AcHSP20-21	Ach13g350901	5.68	21.90	195	Chr13: 14397259.. 14398212（+）	叶绿体Chloroplast
AcHSP20-22	Ach13g383281	6.31	16.61	152	Chr13: 11985660.. 11986118（-）	细胞质Cytoplasmic
AcHSP20-23	Ach13g472651.2	5.94	33.07	297	Chr13: 1497079.. 1498061（-）	叶绿体Chloroplast
AcHSP20-24	Ach14g205801.2	6.87	24.06	216	Chr14: 9422059.. 9423375（+）	叶绿体Chloroplast
AcHSP20-25	Ach14g205811.2	8.86	36.76	329	Chr14: 9424474.. 9428179（+）	叶绿体Chloroplast
AcHSP20-26	Ach15g376651	5.67	22.90	202	Chr15: 4758959.. 4759567（-）	细胞质Cytoplasmic
AcHSP20-27	Ach16g309411.2	7.64	26.14	231	Chr16: 8508810.. 8510240（-）	叶绿体Chloroplast
AcHSP20-28	Ach16g331111	8.83	17.36	153	Chr16: 10507526.. 10508507（+）	细胞质Cytoplasmic
AcHSP20-29	Ach18g165371	4.99	22.60	198	Chr18: 8741760.. 8742825（+）	细胞质Cytoplasmic
AcHSP20-30	Ach19g138461.2	5.08	13.25	114	Chr19: 7272801.. 7273145（+）	细胞质Cytoplasmic
AcHSP20-31	Ach22g015991.2	7.61	26.63	241	Chr22: 8846412.. 8851047（-）	叶绿体Chloroplast
AcHSP20-32	Ach23g121711.2	5.37	22.34	199	Chr23: 4320454.. 4321535（-）	细胞质Cytoplasmic
AcHSP20-33	Ach23g411051.2	9.11	33.06	300	Chr23: 19087591.. 19088582（+）	叶绿体Chloroplast
AcHSP20-34	Ach29g196461.2	4.59	9.84	85	Chr29: 3710817.. 3711074（+）	叶绿体Chloroplast

基因 Gene	基因组登录号 Gene ID	等电点 pI	分子量 Molecular weight/kD	氨基酸数Number of amino acids	染色体定位及基因方向Chromosome localization and gene orientation	亚细胞定位 Subcellular location
AcHSP20-1	Ach00g218421.2	6.57	10.16	88	Chr00: 54153215.. 54154904（-）	细胞质Cytoplasmic
AcHSP20-2	Ach00g239181	7.86	16.14	143	Chr00: 50739806.. 50740562（-）	过氧化物酶体Peroxisomal
AcHSP20-3	Ach00g329621	5.59	23.28	208	Chr00: 90841963.. 90843153（-）	叶绿体Chloroplast
AcHSP20-4	Ach00g479741.2	10.32	9.93	86	Chr00: 135211617.. 135211877（+）	细胞质Cytoplasmic
AcHSP20-5	Ach00g479761.2	9.14	40.18	371	Chr00: 145805825.. 145808080（-）	高尔基体Golgi apparatus
AcHSP20-6	Ach02g036831	7.8	16.56	152	Chr02: 4423490.. 4423948（-）	细胞质Cytoplasmic
AcHSP20-7	Ach02g046741.2	6.21	25.55	225	Chr02: 516872.. 518080（-）	叶绿体Chloroplast
AcHSP20-8	Ach02g223291	8.86	17.61	153	Chr02: 4090770.. 4091231（-）	细胞质Cytoplasmic
AcHSP20-9	Ach03g045661	7.63	25.69	232	Chr03: 6797048.. 6798489（-）	叶绿体Chloroplast
AcHSP20-10	Ach03g451751.2	9.15	11.12	96	Chr03: 1045510.. 1045800（-）	叶绿体Chloroplast
AcHSP20-11	Ach04g161211.2	6.45	24.77	215	Chr04: 6179073.. 6180486（-）	细胞质Cytoplasmic
AcHSP20-12	Ach06g418911.2	6.32	16.60	152	Chr06: 2925504.. 2925962（-）	细胞质Cytoplasmic
AcHSP20-13	Ach10g027441.2	5.4	20.33	181	Chr10: 4989172.. 4989717（-）	细胞质Cytoplasmic
AcHSP20-14	Ach10g027451	7.77	25.72	228	Chr10: 4973842.. 4974528（-）	细胞质Cytoplasmic
AcHSP20-15	Ach10g027461.2	6.34	24.95	221	Chr10: 4984511.. 4985176（+）	叶绿体Chloroplast
AcHSP20-16	Ach10g143161	9.3	25.78	228	Chr10: 11814664.. 11815350（+）	细胞质Cytoplasmic
AcHSP20-17	Ach11g263431.2	5.05	21.72	196	Chr11: 8776984.. 8777574（-）	细胞质Cytoplasmic
AcHSP20-18	Ach11g425231.2	4.43	10.59	90	Chr11: 12263315.. 12263587（-）	细胞质Cytoplasmic
AcHSP20-19	Ach12g058351.2	8.69	17.21	152	Chr12: 6955029.. 6956940（+）	细胞核Nucleus
AcHSP20-20	Ach13g308811	5.94	33.07	297	Chr13: 1116598.. 1117580（+）	叶绿体Chloroplast
AcHSP20-21	Ach13g350901	5.68	21.90	195	Chr13: 14397259.. 14398212（+）	叶绿体Chloroplast
AcHSP20-22	Ach13g383281	6.31	16.61	152	Chr13: 11985660.. 11986118（-）	细胞质Cytoplasmic
AcHSP20-23	Ach13g472651.2	5.94	33.07	297	Chr13: 1497079.. 1498061（-）	叶绿体Chloroplast
AcHSP20-24	Ach14g205801.2	6.87	24.06	216	Chr14: 9422059.. 9423375（+）	叶绿体Chloroplast
AcHSP20-25	Ach14g205811.2	8.86	36.76	329	Chr14: 9424474.. 9428179（+）	叶绿体Chloroplast
AcHSP20-26	Ach15g376651	5.67	22.90	202	Chr15: 4758959.. 4759567（-）	细胞质Cytoplasmic
AcHSP20-27	Ach16g309411.2	7.64	26.14	231	Chr16: 8508810.. 8510240（-）	叶绿体Chloroplast
AcHSP20-28	Ach16g331111	8.83	17.36	153	Chr16: 10507526.. 10508507（+）	细胞质Cytoplasmic
AcHSP20-29	Ach18g165371	4.99	22.60	198	Chr18: 8741760.. 8742825（+）	细胞质Cytoplasmic
AcHSP20-30	Ach19g138461.2	5.08	13.25	114	Chr19: 7272801.. 7273145（+）	细胞质Cytoplasmic
AcHSP20-31	Ach22g015991.2	7.61	26.63	241	Chr22: 8846412.. 8851047（-）	叶绿体Chloroplast
AcHSP20-32	Ach23g121711.2	5.37	22.34	199	Chr23: 4320454.. 4321535（-）	细胞质Cytoplasmic
AcHSP20-33	Ach23g411051.2	9.11	33.06	300	Chr23: 19087591.. 19088582（+）	叶绿体Chloroplast
AcHSP20-34	Ach29g196461.2	4.59	9.84	85	Chr29: 3710817.. 3711074（+）	叶绿体Chloroplast

基因对Gene pairs	Ka	Ks	Ka/Ks
AcHSP20-7/AcHSP20-10	0.231 715 656	0.423 821 440	0.546 729 433
AcHSP20-7/AcHSP20-9	0.209 150 940	0.704 599 321	0.296 836 704
AcHSP20-10/AcHSP20-9	0.322 818 372	0.919 187 813	0.351 199 578
AcHSP20-18/AcHSP20-27	0.102 831 910	0.278 905 623	0.368 697 875
AcHSP20-20/AcHSP20-33	0.104 764 648	0.138 864 084	0.754 440 205
AcHSP20-23/AcHSP20-33	0.104 764 648	0.138 864 084	0.754 440 205
AcHSP20-21/AcHSP20-34	0.092 066 167	0.397 378 202	0.231 683 988
AcHSP20-30/AcHSP20-32	0.239 953 460	0.526 314 021	0.455 913 107