梨矮生相关基因转录组筛选与功能分析

doi:10.16420/j.issn.0513-353x.2024-0481

摘要/Abstract

摘要：

为筛选梨矮生相关基因并分析其功能，以12个株型不同的梨品种（系）为试材，于新梢生长期测定其新梢长度和节间长度；利用转录组测序技术在‘早酥’（普通型）、‘锦香’（半矮生型）和‘中矮1号’（矮生型）梨不同时期生长点、叶片和韧皮部中筛选差异表达基因，并结合在12个品种（系）新梢韧皮部中的表达验证，筛选与矮生性状相关的候选基因，将候选基因PcGA2ox2转化烟草分析其功能。结果表明，‘早酥’‘锦香’和‘中矮1号’的新梢长度在新梢生长初期差异较小，在生长中后期矮生品种短于普通品种；6个普通型品种生长中后期的新梢长度和节间长度均显著长于6个矮生或半矮生型品种（系）；‘早酥’‘锦香’和‘中矮1号’不同时期不同组织不同差异分组两两样品间的差异表达基因数量在1 882 ~ 5 729之间，其中，在叶片和韧皮部中的表达量均随枝条长度缩短而升高或降低的差异表达基因有51个；选择在3个品种新梢韧皮部中差异表达的9个基因利用RT-qPCR在12个品种（系）中进行验证，有4个基因表现出在矮生型品种（系）中的表达显著高于或低于普通型品种（系）的趋势，确定为矮生性状相关候选基因；其中1个矮生候选基因为GA2ox家族基因。进一步在‘中矮1号’基因组中鉴定出了15个GA2ox家族成员，分析发现PcGA2ox2在矮生型品种新梢韧皮部中的表达显著高于普通型品种。将PcGA2ox2在烟草中过表达，转化植株较对照明显变小。综上，认为PcGA2ox2在‘中矮1号’的矮生中起重要作用。

关键词: 梨, 矮生, 转录组分析, 赤霉素, PcGA2ox

Abstract:

To screen the dwarf trait related genes，the shoot length and internode length of 12 pear varieties or selections with different plant architectures were measured during the new shoot growth period. The genes differentially expressed in the growth points，leaves and phloem among three pear varieties ‘Zaosu’（vigorous），‘Jinxiang’（semi-dwarf）and‘Zhongai 1’（dwarf）at different periods were screened by transcripome sequencing analysis. Combined with the expression verification in new shoot phloem of 12 varieties or selections，dwarf trait related candidate genes were obtained. One of the candidate genes PcGA2ox2 was overexpressed in tobacco to analyze its function. The results showed that the new shoot length of‘Zaosu’，‘Jinxiang’and‘Zhongai 1’had a slight difference in the early growth period，while showed a significant difference in the middle and late growth period of new shoot. The internode length of dwarf varieties were shorter than that of vigorous varieties in all test periods. The shoot length and internode length of six vigorous varieties were significantly longer than those of six dwarf or semi-dwarf varieties or selections in the middle and late growth period of new shoot. The number of differentially expressed genes between two samples ranged from 1 882 to 5 729 in different tissues，and periods of three varieties. Among them，51 differentially expressed genes showed an increase or decrease in expression levels with the decrease of new shoot length in both leaf and phloem. Nine differentially expressed genes in new shoot phloem of three varieties were selected to verify the expression levels by RT-qPCR in the phloem of 12 pear varieties or selections in the late growth period of new shoot. Among them，four genes showed a tendency of significant higher or lower expression in dwarf varieties or selections than that in vigorous varieties or selections，and were identified as candidate genes related to dwarf trait. One of the dwarf candidate genes was a member of GA2ox family genes. Total 15 members of the GA2ox gene family were further identified in the‘Zhongai 1’genome，and the expression of the candidate PcGA2ox2 gene was significantly higher in new shoot phloem of dwarf varieties or selections than that of vigorous varieties. Then PcGA2ox2 gene was overexpressed in tobacco，the transformed plants were significantly smaller than the control plants. In conclusion，PcGA2ox2 gene plays a key role in the dwarf trait of‘Zhongai 1’pear.

Key words: pear, dwarf, transcriptome analysis, gibberellin, PcGA2ox

吝茹雪, 王斐, 张艳杰, 马力, 刘肖烽, 李舒然, 刘亚龙, 王晨, 姜淑苓, 欧春青. 梨矮生相关基因转录组筛选与功能分析[J]. 园艺学报, 2025, 52(3): 545-560.

LIN Ruxue, WANG Fei, ZHANG Yanjie, MA Li, LIU Xiaofeng, LI Shuran, LIU Yalong, WANG Chen, JIANG Shuling, OU Chunqing. Transcriptome Screening and Functional Analysis of Dwarf Related Genes in Pear[J]. Acta Horticulturae Sinica, 2025, 52(3): 545-560.

图/表 12

参考文献 51

[1]	Appleford N E, Lenton J R. 1991. Gibberellins and leaf expansion in near-isogenic wheat lines containing Rht1 and Rht3 dwarfing alleles. Planta, 183 (2):229-236. doi: 10.1007/BF00197793 pmid: 24193625
[2]	Chen Xuesen, Wang Nan, Peng Futian, Mao Zhiquan, Yin Chengmiao, Jiang Yuanmao, Ge Shunfeng, Hu Dagang, Li Yuanyuan, Du Yuanpeng, Yao Yuxin, Zhang Zongying. 2024. Advances in quality and maturity breeding of important deciduous fruit trees in China. Acta Horticulturae Sinica, 51 (1):8-26. (in Chinese) doi: 10.16420/j.issn.0513-353x.2023-0271 URL
	陈学森, 王楠, 彭福田, 毛志泉, 尹承苗, 姜远茂, 葛顺峰, 胡大刚, 李媛媛, 杜远鹏, 姚玉新, 张宗营. 2024. 中国重要落叶果树果实品质和熟期育种研究进展. 园艺学报, 51 (1):8-26. doi: 10.16420/j.issn.0513-353x.2023-0271 URL
[3]	Chen Y, Zhang M M, Wang X F, Shao Y, Hu X Y, Cheng J, Zheng X B, Tan B, Ye X, Wang W, Li J D, Li M, Zhang L L, Feng J C. 2023. Peach DELLA protein PpeDGYLA is not degraded in the presence of active GA and causes dwarfism when overexpressed in poplar and Arabidopsis. International Journal of Molecular Sciences, 24 (7):6789.
[4]	Chen Y J, Orlov Y L, Chen M. 2024. Deciphering the molecular mechanism of the intermediate secondary growth and internode elongation of the castor bean(Ricinus communis L.)by the combined analysis of the transcriptome and metabolome. International Journal of Molecular Sciences, 25 (2):1053.
[5]	Cheng Feifei. 2012. Cloning and function analysis of GA2-oxidase gene in dwarf pear of‘Zhongai 1’[M. D. Dissertation]. Xingcheng: Chinese Academy of Agricultural Sciences. (in Chinese)
	程飞飞. 2012. 矮生梨‘中矮1号’GA2-oxidase基因的克隆与功能分析[硕士论文]. 兴城: 中国农业科学院果树研究所.
[6]	Cheng Feifei, Ou Chunqing, Jiang Shuling, Wang Fei, Ma Li, Li Lianwen. 2011. Cloning and expression analysis of ent-kaurene synthase gene in pear. Journal of Shenyang Agricultural University, 42 (6):677-682. (in Chinese)
	程飞飞, 欧春青, 姜淑苓, 王斐, 马力, 李连文. 2011. 梨内根-贝壳杉烯合酶基因克隆及表达分析. 沈阳农业大学学报, 42 (6):677-682.
[7]	Cheng J, Ma J, Zheng X, L H, Zhang M, Tan B, Ye X, Wang W, Zhang L, Li Z, Li J, Feng J. 2021. Functional analysis of the gibberellin 2-oxidase gene family in peach. Frontiers in Plant Science, 12:619158.
[8]	Coll-Garcia D, Mazuch J, Altmann T, Müssig C. 2004. EXORDIUM regulates brassinosteroid-responsive genes. Febs Letters, 563 (1-3):82-86. doi: 10.1016/S0014-5793(04)00255-8 pmid: 15063727
[9]	Cui Z H, Zhang H, Galarneau E R A, Yang Y J, Li D L, Song J K, Ma C H, Zhang S L, Wang R. 2021. Metabolome analysis reveals important compounds related to dwarfing effect of interstock on scion in pear. Annals of Applied Biology, 179 (1):108-122.
[10]	Davidson S E, Smith J J, Helliwell C A, Poole A T, Reid J B. 2004. The pea gene LH encodes ent-kaurene oxidase. Plant Physiology, 134 (3):1123-1134. doi: 10.1104/pp.103.032706 pmid: 14988475
[11]	Dong L L, Yang C X, Wang J, Li J J, Zhao M, Li D L, Qiu Z Y, Ma C H, Cui Z H. 2024. Insights into the dwarfing mechanism of pear(Pyrus betulaefolia)based on anatomical and structural analysis using X-ray scanning. Horticultural Plant Journal, 10 (2):355-366.
[12]	Farrar K, Evans I M, Topping J F, Souter M A, Nielsen J E, Lindsey K. 2003. EXORDIUM-a gene expressed in proliferating cells and with a role in meristem function,identified by promoter trapping in Arabidopsis. The Plant Journal, 33 (1):61-73.
[13]	Feng Y, Zhang X, Wu T, Xu X, Han Z, Wang Y. 2017. Methylation effect on IPT5b gene expression determines cytokinin biosynthesis in apple rootstock. Biochemical and Biophysical Research Communications, 482 (4):604-609. doi: S0006-291X(16)31943-X pmid: 27865843
[14]	Florea L, Song L, Salzberg S L. 2013. Thousands of exon skipping events differentiate among splicing patterns in sixteen human tissues. F1000 Research, 2:188.
[15]	Gao J R, Quan H Y, Yuan L Z, Li Q Y, Qiao L, Li W Q. 2024. Map-based cloning and functional analysis of a new allele of D1,a gene controlling plant height in rice(Oryza sativa L.). Chinese Journal of Rice Science, 38 (2):140-149.
[16]	Gu Wenyi, Wei Haibin. 2021. Selection of dwarfing and densely planted tree form of soft pear,a local characteristic fruit in Qinghai. Science and Technology of Qinghai Agriculture and Forestry,(3):54-58. (in Chinese)
	顾文毅, 魏海斌. 2021. 青海地方特色果品软儿梨矮化密植树形的筛选. 青海农林科技,(3):54-58.
[17]	Hao Ningning, Duan Xuwei, Ou Chunqing, Chen Qiuju, Wang Fei, Jiang Shuling, Ma Li. 2017. Cloning and functional analysis of LUE1 gene from‘Zhongai 1’pear. Acta Horticulturae Sinica, 44 (5):839-849. (in Chinese)
	郝宁宁, 段续伟, 欧春青, 陈秋菊, 王斐, 姜淑苓, 马力. 2017. 梨‘中矮1号’LUE1基因的克隆及功能分析. 园艺学报, 44 (5):839-849. doi: 10.16420/j.issn.0513-353x.2016-0801
[18]	Hedden P, Thomas S G. 2012. Gibberellin biosynthesis and its regulation. Biochemical Journal, 444 (1):11-25. doi: 10.1042/BJ20120245 pmid: 22533671
[19]	Herbst R S. 2004. Review of epidermal growth factor receptor biology. International Journal of Radiation Oncology Biology Physics, 59 (2):S21-S26.
[20]	Hirano K, Kouketu E, Katoh H, Aya K, Ueguchi-Tanaka M, Matsuoka M. 2012. The suppressive function of the rice DELLA protein SLR1 is dependent on its transcriptional activation activity. The Plant Journal, 71 (3):443-453. doi: 10.1111/j.1365-313X.2012.05000.x pmid: 22429711
[21]	Hsieh K T, Chen Y T, Hu T J, Lin S M, Hsieh C H, Liu S H, Shiue S Y, Lo S F, Wang I W, Tseng C S, Chen L J. 2021. Comparisons within the rice GA 2-oxidase gene family revealed three dominant paralogs and a functional attenuated gene that led to the identification of four amino acid variants associated with GA deactivation capability. Rice, 14 (1):70.
[22]	Hu Z Z, Wu Z H, Zhu Q Q, Ma M R, Li Y, Dai X K, Han S P, Xiang S Z, Yang S T, Luo J, Kong Q S, Ding J H. 2024. Multilayer regulatory landscape and new regulators identification for bud dormancy release and bud break in Populus. Plant,Cell & Environment, 47 (8):3181-3197.
[23]	Jiang Shuling, Jia Jingxian, Ji Baosheng, Ma Li. 2000. Pear dwarfing rootstock-Zhongai 1. China Fruits,(3):4-6. (in Chinese)
	姜淑苓, 贾敬贤, 纪宝生, 马力. 2000. 梨矮化砧木——中矮1号. 中国果树,(3):4-6.
[24]	Kim D, Langmead B, Salzberg S L. 2015. HISAT:a fast spliced aligner with low memory requirements. Nature Methods, 12 (4):357-360. doi: 10.1038/NMETH.3317
[25]	Li S, Liu Z, Chen G, Qanmber G, Lu L, Zhang J, Ma S, Yang Z, Li F. 2021. Identification and analysis of GhEXO gene family indicated that GhEXO7_At promotes plant growth and development through brassinosteroid signaling in cotton(Gossypium hirsutum L.). Frontiers in Plant Science, 12:719889.
[26]	Li Wenyuan, Lin Menghua, Li Yahui, Yu Quanqi, Liang Ying, Zhang Zhiyong. 2024. Advances in metabolomics of pear fruit. Acta Horticulturae Sinica, 51 (7):1595-1609. (in Chinese) doi: 10.16420/j.issn.0513-353x.2023-0699 URL
	李文远, 林梦桦, 李亚辉, 于全琦, 梁颖, 张志勇. 2024. 梨果实代谢组学研究进展. 园艺学报, 51 (7):1595-1609.
[27]	Liang Z, Wang Q, Sun M, Du R, Jin W, Liu S. 2024. Transcriptome and metabolome profiling reveal the effects of hormones on current-year shoot growth in Chinese‘Cuiguan’pear grafted onto vigorous rootstock‘Duli’and dwarf rootstock‘Quince A’. BMC Plant Biology, 24 (1):169.
[28]	Lin J, Feng D, Li Y, Xiao S, Xuan W. 2023. Comparing the structural characteristics and expression of GA2ox gene in dwarf banana and its wild type. Journal of Plant Biochemistry and Biotechnology, 32 (1):76-84.
[29]	Liu X, Wang J, Sabir I A, Sun W, Wang L, Xu Y, Zhang N, Liu H, Jiu S, Liu L, Zhang C. 2022. PavGA2ox-2L inhibits the plant growth and development interacting with PavDWARF in sweet cherry(Prunus avium L.). Plant Physiology and Biochemistry, 186:299-309.
[30]	Luo Y, Dong X, Yu T, Shi X, Li Z, Yang W, Widmer A, Karrenberg S. 2015. A single nucleotide deletion in Gibberellin20-oxidase1 causes alpine dwarfism in Arabidopsis. Plant Physiology, 168 (3):930-937.
[31]	Ou C Q, Wang F, Wang J H, Li S, Zhang Y J, Fang M, Ma L, Zhao Y N, Jiang S L. 2019. A de novo genome assembly of the dwarfing pear rootstock Zhongai 1. Scientific Data, 6 (1):281.
[32]	Ou Chunqing, Jiang Shuling, Wang Fei, Wang Zhigang, Ma li, Li Lianwen. 2013. Cloning and expression analysis of ent-kaurenoic acid oxidase gene(PcKAO1)in pear. Acta Horticulturae Sinica, 40 (5):849-858. (in Chinese)
	欧春青, 姜淑苓, 王斐, 王志刚, 马力, 李连文. 2013. 梨贝壳杉烯酸氧化酶基因PcKAO1的克隆与表达分析. 园艺学报, 40 (5):849-858.
[33]	Peng J R, Carol P, Richards D E, King K E, Cowling R J, Murphy G P, Harberd N P. 1997. The Arabidopsis GAI gene defines a signaling pathway that negatively regulates gibberellin responses. Genes & Development, 11 (23):3194-3205.
[34]	Salcini A E, Chen H, Iannolo G, De Camilli P, Di Fiore P P. 1999. Epidermal growth factor pathway substrate 15,Eps15. International Journal of Biochemistry & Cell Biology, 31 (8):805-809.
[35]	Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush G S, Kitano H, Matsuoka M. 2002. Green revolution:a mutant gibberellin-synthesis gene in rice - new insight into the rice variant that helped to avert famine over thirty years ago. Nature, 416 (6882):701-702.
[36]	Scheepers M, Spielmann J, Boulanger M, Carnol M, Bosman B, De Pauw E, Goormaghtigh E, Motte P, Hanikenne M. 2020. Intertwined metal homeostasis,oxidative and biotic stress responses in the Arabidopsis frd3 mutant. The Plant Journal, 102 (1):34-52. doi: 10.1111/tpj.14610 pmid: 31721347
[37]	Schröder F, Lisso J, Lange P, Müssig C. 2009. The extracellular EXO protein mediates cell expansion in Arabidopsis leaves. BMC Plant Biology, 9 (20):1-12.
[38]	Sha Guangli. 2007. Apomictic dwarf apple germplasm innovation[Ph. D. Dissertation]. Tai’an: Shandong Agricultural University. (in Chinese)
	沙广利. 2007. 苹果无融合生殖矮化种质创新[博士论文]. 泰安: 山东农业大学.
[39]	Singh D, Attri B K, Gill R K, Bariwal J. 2016. Review on EGFR inhibitors:critical updates. Mini-Reviews in Medicinal Chemistry, 16 (14):1134-1166.
[40]	Tang Changyong. 2015. Screening of dwarf genes and clone and expression analysis of PcAHS gene from‘Zhongai 1’pear[M. D. Dissertation]. Xingcheng: Chinese Academy of Agricultural Sciences. (in Chinese)
	汤常永. 2015. 梨‘中矮1号’矮生基因的筛选及PcAHS克隆与表达分析[硕士论文]. 兴城: 中国农业科学院.
[41]	Tang L, Xu H, Wang Y, Wang H, Li Z, Liu X, Shu Y, Li G, Liu W, Ying J, Tong X, Yao J, Xiao W, Tang S, Ni S, Zhang J. 2021. OsABF1 represses Gibberellin biosynthesis to regulate plant height and seed germination in Rice (Oryza sativa L.). International Journal of Molecular Sciences, 22 (22):12220.
[42]	Teshome S, Kebede M. 2021. Analysis of regulatory elements in GA2ox,GA3ox and GA20ox gene families in Arabidopsis thaliana:an important trait. Biotechnology & Biotechnological Equipment, 35 (1):1603-1612.
[43]	Wang L, Feng Z, Wang X, Wang X, Zhang X. 2010. DEGseq:an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics, 26 (1):136-138.
[44]	Wang Y, Deng D. 2014. Molecular basis and evolutionary pattern of GA-GID1-DELLA regulatory module. Molecular Genetics and Genomics, 289 (1):1-9. doi: 10.1007/s00438-013-0797-x pmid: 24322346
[45]	Winkler R G, Helentjaris T. 1995. The maize Dwarf3 gene encodes a cytochrome P450-mediated early step in gibberellin biosynthesis. The Plant Cell, 7 (8):1307-1317.
[46]	Yan J D, Xiang F J, Yang P, Li X, Zhong M, He R Q, Li X M, Peng W S, Liu X M, Zhao X Y. 2021. Overexpression of BnGA2ox2,a rapeseed gibberellin 2-oxidase,causes dwarfism and increased chlorophyll and anthocyanin accumulation in Arabidopsis and rapeseed. Plant Growth Regulation, 93 (1):65-77.
[47]	Yang G, Sun M, Brewer L, Tang Z, Nieuwenhuizen N, Cooney J, Xu S, Sheng J, Andre C, Xue C, Rebstock R, Yang B, Chang W, Liu Y, Li J, Wang R, Qin M, Brendolise C, Allan A C, Espley R V, Lin-Wang K, Wu J. 2024. Allelic variation of BBX24 is a dominant determinant controlling red coloration and dwarfism in pear. Plant Biotechnology Journal, 22 (6):1468-1490.
[48]	Ye Yu, Ou Chunqing, Wang Fei, Zhang Yanjie, Ma Li, Jiang Shuling. 2022. Progress in research on the dwarfing mechanism of pear dwarfing root stocks. Journal of Fruit Science, 39 (11):2163-2171. (in Chinese)
	叶宇, 欧春青, 王斐, 张艳杰, 马力, 姜淑苓. 2022. 梨矮化砧木致矮机制研究进展. 果树学报, 39 (11):2163-2171.
[49]	Zhang Yuliang, Zong Yu, Song Jiankun, Li Dingli, Wang Xinhui, Wang Ran. 2017. Seedling growth of four pear rootstocks and changes in mineral elements in their leaves,shoots and roots during growing season. Journal of Fruit Science, 34 (12):1545-1555. (in Chinese)
	张玉良, 宗宇, 宋建坤, 李鼎立, 王新慧, 王然. 2017. 4种梨砧木实生苗生长及其叶片、新梢和根中的矿质元素含量的变化. 果树学报, 34 (12):1545-1555.
[50]	Zhao H, Wan S, Huang Y, Li X, Jiao T, Zhang Z, Ma B, Zhu L, Ma F, Li M. 2023. The transcription factor MdBPC 2 alters apple growth and promotes dwarfing by regulating auxin biosynthesis. The Plant Cell, 36 (3):585-604.
[51]	Zheng X, Zhang H, Xiao Y, Wang C, Tian Y. 2019. Deletion in the promoter of PcPIN-L affects the polar auxin transport in dwarf pear(Pyrus communis L.). Scientific Reports, 9 (1):18645.

基因ID Gene ID	正向引物（5′-3′）Forward primer	反向引物（5′-3′）Reverse primer
Pdr15g002180	CCCCTTGTGGCTCCCAATA	CCTGAAAGTATCCATTCCCAAAT
Pdr3g011110	TGCTCCATGTCCATTTCCTTC	ACTCGGTGTTTGATGCTCTTGTAT
New Gene_2402	GATCTCTTCCCTTATTTTTCATCAGTT	GTTCCTACCGTGCCCTTTGT
Pdr11g021900	CTCGGGTTTCTACTTCTACACTGG	AGGGCACAATTCACTGCTCTTA
Pdr14g004080	TGGCGGTTCGGCTTGG	GGGTTGACCATTGGATTGTCTC
Pdr13g000360	CAAAACGGAACACGCAAACC	TTACAAGGAAGCACAAGAGGACC
Pdr10g017040	AGGTTGTTGAAAATGGAGCAGC	GAGGCACCTGTGAAAGTTGTCC
Pdr17g005180	CTCCCAACTTTCGCCGTCT	AAAAAGGCTCGGTCCAGAAAT
Pdr7g012000	TCCGAAGATGTTGGGGACC	AACTGCTCACATGGCACGGT
PcActin （GU830958.1）	GGAATGGAAGCTGCTGGC	CGTCATACTCACCCTTGGAAATC
NtActin（NM_001325686.1）	CTACTTACTGAA GCACCCTTGAATCC	CCTGCCCA TCTGGTAACTCATAGC

基因ID Gene ID	正向引物（5′-3′）Forward primer	反向引物（5′-3′）Reverse primer
Pdr15g002180	CCCCTTGTGGCTCCCAATA	CCTGAAAGTATCCATTCCCAAAT
Pdr3g011110	TGCTCCATGTCCATTTCCTTC	ACTCGGTGTTTGATGCTCTTGTAT
New Gene_2402	GATCTCTTCCCTTATTTTTCATCAGTT	GTTCCTACCGTGCCCTTTGT
Pdr11g021900	CTCGGGTTTCTACTTCTACACTGG	AGGGCACAATTCACTGCTCTTA
Pdr14g004080	TGGCGGTTCGGCTTGG	GGGTTGACCATTGGATTGTCTC
Pdr13g000360	CAAAACGGAACACGCAAACC	TTACAAGGAAGCACAAGAGGACC
Pdr10g017040	AGGTTGTTGAAAATGGAGCAGC	GAGGCACCTGTGAAAGTTGTCC
Pdr17g005180	CTCCCAACTTTCGCCGTCT	AAAAAGGCTCGGTCCAGAAAT
Pdr7g012000	TCCGAAGATGTTGGGGACC	AACTGCTCACATGGCACGGT
PcActin （GU830958.1）	GGAATGGAAGCTGCTGGC	CGTCATACTCACCCTTGGAAATC
NtActin（NM_001325686.1）	CTACTTACTGAA GCACCCTTGAATCC	CCTGCCCA TCTGGTAACTCATAGC

编号 Code	采样时间/ （M-D） Sampling time	组织 Tissue	比对材料 Comparative materials	高表达 High expression	低表达 Low expression	合计 Total
1	04-28	生长点 Growing point	早酥 vs. 锦香 Zaosu vs. Jinxiang	1 723	1 244	2 967
2			早酥vs. 中矮1号Zaosu vs. Zhongai 1	1 158	724	1 882
3			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	1 269	1 111	2 380
			1 ~ 3合计 Total of 1-3	3 301	2 671	4 865
4	05-18	叶片Leaf	早酥 vs. 锦香 Zaosu vs. Jinxiang	2 503	2 244	4 747
5			早酥vs. 中矮1号Zaosu vs. Zhongai 1	3 450	2 049	5 499
6			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	2 482	1 292	3 774
7		韧皮部Phloem	早酥 vs. 锦香 Zaosu vs. Jinxiang	2 679	2 181	4 860
8			早酥vs. 中矮1号Zaosu vs. Zhongai 1	2 540	1 717	4 257
9			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	1 463	1 069	2 532
			4 ~ 9合计 Total of 4-9	7 878	6 252	11 741
10	06-15	叶片Leaf	早酥 vs. 锦香 Zaosu vs. Jinxiang	2 890	1 859	4 749
11			早酥vs. 中矮1号Zaosu vs. Zhongai 1	2 951	2 333	5 284
12			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	2 139	2 562	4 701
13		韧皮部Phloem	早酥 vs. 锦香 Zaosu vs. Jinxiang	3 043	2 686	5 729
14			早酥vs. 中矮1号Zaosu vs. Zhongai 1	2 606	1 956	4 562
15			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	1 596	1 167	2 763
			10 ~ 15合计 Total of 10-15	7 907	7 275	12 201
16	04-28	生长点 Growing point	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	6	3	9
17	05-18	叶片Leaf	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	124	72	196
18		韧皮部Phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	37	43	80
19	06-15	叶片Leaf	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	113	77	190
20		韧皮部Phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	51	46	97
			16 ~ 20合计 Total of 16-20	276	190	466
21	05-18， 06-15	叶片Leaf	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	18	20	38
22		韧皮部Phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	11	10	21
			21 ~ 22合计 Total of 21-22	26	25	51
23	05-18， 06-15	叶片和韧皮部 Leaf and phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	3	5	8

编号 Code	采样时间/ （M-D） Sampling time	组织 Tissue	比对材料 Comparative materials	高表达 High expression	低表达 Low expression	合计 Total
1	04-28	生长点 Growing point	早酥 vs. 锦香 Zaosu vs. Jinxiang	1 723	1 244	2 967
2			早酥vs. 中矮1号Zaosu vs. Zhongai 1	1 158	724	1 882
3			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	1 269	1 111	2 380
			1 ~ 3合计 Total of 1-3	3 301	2 671	4 865
4	05-18	叶片Leaf	早酥 vs. 锦香 Zaosu vs. Jinxiang	2 503	2 244	4 747
5			早酥vs. 中矮1号Zaosu vs. Zhongai 1	3 450	2 049	5 499
6			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	2 482	1 292	3 774
7		韧皮部Phloem	早酥 vs. 锦香 Zaosu vs. Jinxiang	2 679	2 181	4 860
8			早酥vs. 中矮1号Zaosu vs. Zhongai 1	2 540	1 717	4 257
9			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	1 463	1 069	2 532
			4 ~ 9合计 Total of 4-9	7 878	6 252	11 741
10	06-15	叶片Leaf	早酥 vs. 锦香 Zaosu vs. Jinxiang	2 890	1 859	4 749
11			早酥vs. 中矮1号Zaosu vs. Zhongai 1	2 951	2 333	5 284
12			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	2 139	2 562	4 701
13		韧皮部Phloem	早酥 vs. 锦香 Zaosu vs. Jinxiang	3 043	2 686	5 729
14			早酥vs. 中矮1号Zaosu vs. Zhongai 1	2 606	1 956	4 562
15			锦香vs. 中矮1号Jinxiang vs. Zhongai 1	1 596	1 167	2 763
			10 ~ 15合计 Total of 10-15	7 907	7 275	12 201
16	04-28	生长点 Growing point	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	6	3	9
17	05-18	叶片Leaf	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	124	72	196
18		韧皮部Phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	37	43	80
19	06-15	叶片Leaf	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	113	77	190
20		韧皮部Phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	51	46	97
			16 ~ 20合计 Total of 16-20	276	190	466
21	05-18， 06-15	叶片Leaf	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	18	20	38
22		韧皮部Phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	11	10	21
			21 ~ 22合计 Total of 21-22	26	25	51
23	05-18， 06-15	叶片和韧皮部 Leaf and phloem	早酥 vs. 锦香vs. 中矮1号Zaosu vs. Jinxiang vs. Zhongai 1	3	5	8

组织 Tissue	随新梢长度的缩短表达升高的基因 Genes that increase in expression with the decrease of new shoot length		随新梢长度的缩短表达降低的基因 Genes that decrease in expression with the decrease of new shoot length
组织 Tissue	ID	注释Annotation	ID	注释Annotation
叶片 Leaf	Pdr8g014740	Probable disease resistance protein At4g27220	Pdr16g015850	Protein SIEVE ELEMENT OCCLUSION B-like
		isoform X2	Pdr0g057320	Uncharacterized protein
	Pdr5g018850	G-type lectin S-receptor-like serine/threonine-protein kinase At4g27290 isoform X1	Pdr11g012240	Palmitoyl-monogalactosyldiacylglycerol delta-7 desaturase，chloroplastic-like
	Pdr11g021340	UDP-glycosyltransferase 87A1-like	Pdr17g003340	UDP-glycosyltransferase 83A1-like
	Pdr10g027130	Carbonic anhydrase 2-like isoform X1	Pdr17g001240	Putative glucose-6-phosphate 1-epimerase isoform X1
	Pdr7g014190	3-ketoacyl-CoA synthase 19-like	Pdr10g015920	Putative amidase C869.01 isoform X1
	Pdr0g030210	Probable serine/threonine-protein kinase	Pdr11g022920	Uncharacterized protein
		DDB_G0282963	Pdr16g009800	Geraniol 8-hydroxylase-like isoform X1
	Pdr11g002090	LEC14B protein-like isoform X1	Pdr0g061140	Succinate dehydrogenase subunit 5，mitochondrial-like
	Pdr9g003550	Uncharacterized protein At1g03900-like	Pdr11g004540	Probable leucine-rich repeat receptor-like
	Pdr4g000300	Serine carboxypeptidase-like 20		serine/threonine-protein kinase
	Pdr6g013710	-	Pdr9g003650	Uncharacterized protein
	Pdr10g000140	Early nodulin-like protein 2	Pdr9g006840	Acid phosphatase 1-like
	Pdr2g011040	TMV resistance protein N-like	Pdr3g014200	Uncharacterized protein
	NewGene_1645	-	Pdr9g000780	Disease resistance protein At4g27190-like isoform X2
	NewGene_492	Uncharacterized protein	NewGene_3128	40S ribosomal protein S29
	NewGene_7522	Uncharacterized protein
韧皮部	Pdr15g002180	Protein EXORDIUM-like	Pdr6g000360	Cinnamoyl-CoA reductase 2-like
Phloem	Pdr0g064490	Uncharacterized protein	Pdr16g022720	Uncharacterized protein
	Pdr9g009480	Uncharacterized protein	Pdr1g005260	Trans-resveratrol di-O-methyltransferase-like
	Pdr11g021900	Putative disease resistance protein RGA1 isoform	Pdr15g014540	BURP domain protein RD22-like
		X1	Pdr10g006620	CSC1-like protein ERD4
	Pdr1g005700	（+）-neomenthol dehydrogenase-like isoform X1
	Pdr10g000410	Ankyrin repeat-containing protein NPR4-like
	Pdr2g010870	Uncharacterized protein
	Pdr9g000720	DNA polymerase delta catalytic subunit
叶片和韧皮部 Leaf and phloem	Pdr9g008130	Ras-associated and pleckstrin homology domains-containing protein 1-like	Pdr16g008560	Non-functional NADPH-dependent codeinone reductase 2-like
	NewGene_2402	Topless-related protein 1-like isoform X1	Pdr11g007050	Putative disease resistance RPP13-like protein 1
	NewGene_4126	-	Pdr0g010770	Uncharacterized protein
			Pdr7g019110	TMV resistance protein N-like
			NewGene_4125	Uncharacterized protein