Identification of F1 Hybrids and Analysis of Fruit Shape Difference Between Fingered Citron and Citron

doi:10.16420/j.issn.0513-353x.2022-0745

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (7): 1402-1418.doi: 10.16420/j.issn.0513-353x.2022-0745

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Identification of F₁ Hybrids and Analysis of Fruit Shape Difference Between Fingered Citron and Citron

MEI Yulin¹^,², XU Jinjian¹, JIANG Mengyue¹, YU Junhai¹, ZONG Yu¹, CHEN Wenrong¹, LIAO Fanglei¹^,²^,^*(), GUO Weidong¹^,²^,^*()

¹ College of Chemistry and Life Sciences，Zhejiang Normal University，Jinhua，Zhejiang 321004，China
² Zhejiang Provincial Key Laboratory of Biotechnology on Specialty Economic Plants，Jinhua，Zhejiang 321004，China

Received:2022-11-21 Revised:2023-01-18 Published:2023-07-26
Contact: LIAO Fanglei, GUO Weidong

Abstract

Abstract:

In this study，19 F₁ hybrids derived from the cross between Citrus medica L. and C. medica var. L. sarcodactylis Swingle were cultured as materials. SSR loci were selected from the transcriptome data of fingered citron and citron. Based on this，TP-M13-SSR primers were designed and then 19 F₁ hybrids were genotyped. A total of 17 to 32 alleles were obtained from 16 SSR primers，and the average number of alleles was 24.188. The heterozygosity was ranged from 0.188 to 1.000，and the polymorphism index was about 0.944. The results showed that 17 out of 19 F₁ lines were true hybrids，and the hybridization rate was 89.47%. Those materials were phenotyped by morphological photography analysis and the ratio of finger fruit to hesperidium is 2︰3. Paraffin section analysis of early flower buds of fingered citron and citron found the difference between their fruit shape initiating from the carpel bulges since the visible bud stage to the petals emerging stage. From the visible bud stage to 3 d after anthesis of fingered citron and citron，the correlation analysis between the transcriptome data and carpel feature was done，and a number of genes associated with gynoecium formation were identified. These genes were validated in the pistils of citron，fingered citron，and hybrid F₁ lines during the pre-anthesis stage. The results showed that the expression of WOX1 was related to the character of finger fruit from the visible bud stage to the petals emerging stage. The YABBY family coordinated with the classes C，E genes of the ABC model of flower development and WOX family to regulate the fruit shaping in fingered citron.

Key words: citron, fingered citron, SSR, finger fruit, gynoecium formation, WOX1

MEI Yulin, XU Jinjian, JIANG Mengyue, YU Junhai, ZONG Yu, CHEN Wenrong, LIAO Fanglei, GUO Weidong. Identification of F₁ Hybrids and Analysis of Fruit Shape Difference Between Fingered Citron and Citron[J]. Acta Horticulturae Sinica, 2023, 50(7): 1402-1418.

Figures/Tables 13

References 48

[1]	Bowman J L, Smyth D R. 1999. CRABS CLAW, a gene that regulates carpel and nectary development in Arabidopsis, encodes a novel protein with zinc finger and helix-loop-helix domains. Development, 126:2387-2396. doi: 10.1242/dev.126.11.2387 pmid: 10225998
[2]	Chen Min-guan. 2014. Preliminary studies on fruit set and fruit shaped development of fingered citron[M. D. Dissertation]. Jinhua: Zhejiang Normal University. (in Chinese)
	陈民管. 2014. 佛手坐果及果形发育的初步研究[硕士论文]. 金华: 浙江师范大学.
[3]	Chen Q, Atkinson A, Otsuga D, Christensen T, Reynolds L, Drews G N. 1999. The Arabidopsis FLAMENTOUS FLOWER gene is required for flower formation. Development, 126:2715-2726. doi: 10.1242/dev.126.12.2715 pmid: 10331982
[4]	Chen Xiao-lin, Wang Nian, Zhu Yan-lin. 2012. Progress in study of plant atavisms. Acta Agriculturae Shanghai, 28 (1):102-105. (in Chinese)
	陈小灵, 王念, 朱延林. 2012. 植物返祖现象研究. 上海农业学报, 28 (1):102-105.
[5]	Cristina F, Chloé F, Nathanael P, Charlie P S, Eva S, Christophe T, Aurélie C M, Vialette G. 2010. Carpel development. Advances in Botanical Research, 55:1-73.
[6]	Deng H H, Cai Z X, Xiang S Q, Guo Q G, Huang W, Liang G L. 2019. Karyotype analysis of diploid and spontaneously occurring tetraploid blood orange [Citrus sinensis (L.) Osbeck] using multicolor FISH with repetitive DNA sequences as probes. Front in Plant Science, 10:331. doi: 10.3389/fpls.2019.00331 URL
[7]	Dong Qing-hua, Wang Xi-cheng, Zhao Mi-zheng, Song Chang-nian, Ge An-jing, Wang Jing. 2011. Development of EST-derived SSR markers and their application in strawberry genetic diversity analysis. Agricultural Sciences in China, 44 (17):3603-3612. (in Chinese)
	董清华, 王西成, 赵密珍, 宋长年, 葛安静, 王静. 2011. 草莓EST-SSR标记开发及在品种遗传多样性分析中的应用. 中国农业科学, 44 (17):3603-3612. doi: 10.3864/j.issn.0578-1752.2011.17.013
[8]	Drews G N, Bowman J L, Meyerowitz E M. 1991. Negative regulation of the Arabidopsis homeotic gene AGAMOUS by the APETALA2product. Cell, 65:991-1002. doi: 10.1016/0092-8674(91)90551-9 pmid: 1675158
[9]	Editorial Committee of flora of China,Chinese Academy of Sciences. 1997. Flora of China. 43 (2). Beijing:Science Press:184. (in Chinese)
	中国科学院中国植物志编辑委员会. 1997. 中国植物志. 43 (2). 北京:科学出版社:184.
[10]	Fang Jing-gui, Zhang Zhen, Ma Zheng-qiang, Liu Da-jun, Wang San-hong, Lavi U. 2000. The polymorphism and segregation patterns of AFLP markers in the F_l progenies from the cross of two mango cultivars. Scientia Agricultura Sinica, 33 (3):19-24. (in Chinese)
	房经贵, 章镇, 马正强, 刘大钧, 王三红, Lavi U. 2000. AFLP标记在两个芒果品种间杂交Fl代的多态性及分离方式. 中国农业科学, 33 (3):19-24. doi: 10.3864/j.issn.0578-1752.2000-33-3-22-27
[11]	Fang Qian, Zhang Yuan-yuan, Yang Yu-ting, Huang Miao-miao, Fu Qiao-li, Zhou Hui-sha, Chen Weng-rong, Zong Yu, Guo Wei-dong. 2018. SSR mining and polymorphism analysis in leaf transcriptome of blueberry. Acta Horticulturae Sinica, 45 (7):1359-1370. (in Chinese) doi: 10.16420/j.issn.0513-353x.2017-0675
	方茜, 张园园, 杨钰婷, 黄苗苗, 符巧丽, 周慧莎, 陈文荣, 宗宇, 郭卫东. 2018. 越橘叶片转录组SSR发掘及其多态性研究. 园艺学报, 45 (7):1359-1370. doi: 10.16420/j.issn.0513-353x.2017-0675
[12]	Fang Zhi-wei, Li Lun. 2018. Research progress of SSR typing technology. Biological Chemical Engineering, 4 (1):118-121. (in Chinese)
	方治伟, 李论. 2018. SSR分型技术研究进展. 生物化工, 4 (1):118-121.
[13]	Gómez M C, de Folter S, Costa M M, Angenent G C, Sablowski R. 2005. Transcriptional program controlled by the floral homeotic gene AGAMOUS during early organogenesis. Development, 132:429-438. doi: 10.1242/dev.01600 URL
[14]	Guo Ji-chun, Ye Nai-xing, He Xiao-yan. 2004. Genetic variation in the leaf-expansion period of the first hybrid generation tea plants. Journal of Tea Science,(4):255-259. (in Chinese)
	郭吉春, 叶乃兴, 何孝延. 2004. 茶树杂交一代展叶期的遗传变异. 茶叶科学,(4):255-259.
[15]	Han Guo-hui, Xiang Su-qiong, Wang Wei-xing, Wei Xu, He Bo, Li Xiao-lin, Liang Guo-lu. 2010. Identification and genetic diversity of hybrid progenies from Shatian pummelo by SSR. Scientia Agricultura Sinica, 43 (22):4678-4686. (in Chinese)
	韩国辉, 向素琼, 汪卫星, 魏旭, 何波, 李晓林, 梁国鲁. 2010. 沙田柚杂交后代群体的 SSR 鉴定与遗传多样性分析. 中国农业科学, 43 (22):4678-4686.
[16]	Han Xiao-xia. 2017. Function analysis of them YABBY5 and CRC gene in fruit shaping of the fingered citron[M. D. Dissertation]. Jinhua: Zhejiang Normal University. (in Chinese)
	韩晓霞. 2017. YABBY5和CRC基因在佛手果实发育中的功能初探[硕士论文]. 金华: 浙江师范大学.
[17]	Hempel F D, Weigel D, Mandel M A, Ditta G, Zambryski P C, Feldman L J, Yanofsky M F. 1997. Floral determination and expression of floral regulatory genes in Arabidopsis. Development, 124:3845-3853. doi: 10.1242/dev.124.19.3845 pmid: 9367440
[18]	Hu Wen-shun, Huang Ai-ping, Jiang Fan, Jiang Ji, Chen Xiu-ping, Zheng Shao-quan. 2015. Identification and genetic diversity of reciprocal hybrids in longyan (Dimocarpus longan) by SSR. Acta Horticulturae Sinica, 42 (10):1899-1908. (in Chinese)
	胡文舜, 黄爱萍, 姜帆, 蒋际, 陈秀萍, 郑少泉. 2015. 龙眼正反交后代的SSR鉴定及遗传多样性分析. 园艺学报, 42 (10):1899-1908. doi: 10.16420/j.issn.0513-353x.2015-0131
[19]	Jiang D, Zhong G Y, Hong Q B. 2006. Analysis of microsatellites in Citrus unigenes. Acta Genetica Sinica, 33 (4):345-353. pmid: 16625833
[20]	Jones A G, Small C M, Paczolt K A, Ratterman N L. 2010. A practical guide to methods of parentage analysis. Molecular Ecology Resources, 10 (1):6-30. doi: 10.1111/j.1755-0998.2009.02778.x pmid: 21564987
[21]	Kumpatla S P, Mukhopadhyay S. 2005. Mining and survey of simple sequence repeats in expressed sequence tags of dicotyledonous species. Genome, 48 (6):985-998. pmid: 16391668
[22]	Lee J Y, Baum S F, Alvarez J, Patel A, Chitwood D H, Bowman J L. 2005. Activation of CRABS CLAW in the nectaries and carpels of Arabidopsis. The Plant Cell, 17 (1):25-36. doi: 10.1105/tpc.104.026666 URL
[23]	Li Hui-yong, Wang Tian-yu, Li Yu, Shi Yun-su, Song Yan-chun, Lu Ping. 2005. Application of the TP-M13 automated fluorescent-lablled system of SSR genotyping in sorghum. Journal of Plant Genetic Resources, 6 (1):68-70. (in Chinese)
	李会勇, 王天宇, 黎裕, 石云素, 宋艳春, 陆平. 2005. TP-M13自动荧光检测法在高粱SSR基因型鉴定中的应用. 物遗传资源学报, 6 (1):68-70.
[24]	Li Xiang, Yang Nan, Zhao Kai-ge, Chen Yu-xing, Tang Rui-jun, Chen Long-qing. 2013. Development and primer selection of EST-SSR molecular markers based on transcriptome sequencing of Chimonanthus praecox. Journal of Beijing Forestry University, 35 (1):25-32. (in Chinese)
	李响, 杨楠, 赵凯歌, 陈玉星, 唐锐君, 陈龙清. 2013. 蜡梅转录组EST-SSR标记开发与引物筛选. 北京林业大学学报, 35 (1):25-32.
[25]	Liang X Q, Chen X P, Hong Y B, Liu H Y, Zhou G Y, Li S X, Guo B Z. 2009. Utility of EST-derived SSR in cultivated peanut (Arachis hypogaea L.) and Arachis wild species. BMC Plant Biology, 9:35. doi: 10.1186/1471-2229-9-35 URL
[26]	Liao F L, Wang Y J, Chen M G, Chen W R, Zong Y, Li Y Q, Yang L, Han X X, Guo W D. 2017. Low transcription of CmsIAA9 in the basal pistil is related to parthenocarpic fruiting of fingered citron(Foshou). Molecular Breeding, 37 (8):101. doi: 10.1007/s11032-017-0693-x URL
[27]	Liao Fang-lei, Chen Min-guan, Sang Dan, Chen Wen-rong, Guo Wei-dong. 2013. Genetic diversity analysis of fingered citron germplasm by ISSR. Acta Horticulturae Sinica, 40 (11):2222-2228. (in Chinese)
	廖芳蕾, 陈民管, 桑丹, 陈文荣, 郭卫东. 2013. 佛手种质资源遗传多样性的ISSR 分析. 园艺学报, 40 (11):2222-2228.
[28]	Liao Fanglei, Chen Zeyu, Xu Qiyue, Yang Li, Chen Wenrong, Guo Weidong. 2018. A review on fruit-shaping genes study:reference for fruit pattern formation in fingered citron. Acta Horticulturae Sinica, 45 (9):1701-1714. (in Chinese) doi: 10.16420/j.issn.0513-353x.2018-0502
	廖芳蕾, 陈泽宇, 徐启越, 杨莉, 陈文荣, 郭卫东. 2018. 果形建成基因研究进展及其对佛手果形发育研究的启示. 园艺学报, 45 (9):1701-1714. doi: 10.16420/j.issn.0513-353x.2018-0502
[29]	Liao Fang-lei, Han Xiao-xia, Chen Wen-rong, Guo Yan, Zhang Chen-xiao, Chen Ze-yu, Zhou Ya-yan, Guo Wei-dong. 2016. Fruit morphogenesis observation and expression analysis of fruit-shaping related genes in fingered citron. Acta Horticulturae Sinica, 43 (11):2141-2150. (in Chinese) doi: 10.16420/j.issn.0513-353x.2016-0358
	廖芳蕾, 韩晓霞, 陈文荣, 郭艳, 张晨晓, 陈泽宇, 周亚艳, 郭卫东. 2016. 佛手果形发育观察及果形相关基因表达分析. 园艺学报, 43 (11):2141-2150.
[30]	Liu Hang-xiu, Fen Di, Long Chun-rui, Zhou Xiao-yan, Liu Hong-ming, Yang Hong-xia, Du Yu-xia, Guo Li-na, Fu Xiao-meng, Ma Zhao-cheng, Yue Jian-qiang. 2021. Fruit variation and geographical distribution of citron. China Journal of Chinese Materia Medica, 46 (23):6289-6293. (in Chinese) doi: 10.19540/j.cnki.cjcmm.20211012.102 pmid: 34951256
	刘航秀, 冯迪, 龙春瑞, 周先艳, 刘红明, 杨虹霞, 杜玉霞, 郭丽娜, 付小猛, 马兆成, 岳建强. 2021. 枸橼药用植物果实变异及地理分布研究. 中国中药杂志, 46 (23):6289-6293. pmid: 34951256
[31]	Liu Sheng-rui. 2016. High-density genetic map construction and identification of QTLs controlling deciduous trait in Citrus[Ph. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	刘升锐. 2016. 柑橘高密度遗传连锁图谱的构建及落叶性状的QTL定位[博士论文]. 武汉: 华中农业大学.
[32]	Luo Qiu, Zhang Qing-xiang. 2020. Application of SSR technology in fruit trees. Jiangxi Agriculture,(10):36-38. (in Chinese)
	罗球, 张庆祥. 2020. SSR技术在果树上的应用. 江西农业,(10):36-38.
[33]	ÓMaoiléidigh D S, Wuest S E, Rae L, Raganelli A, Ryan P T, Kwasniewska K, Das P, Lohan A J, Loftus B, Graciet E, Wellmer F. 2013. Control of reproductive floral organ identity specification in Arabidopsis by the function regulator AGAMOUS. The Plant Cell, 25 (7):2482-2503. doi: 10.1105/tpc.113.113209 pmid: 23821642
[34]	Schuelke M. 2000. An economic method for the fluorescent labeling of PCR fragments. Nature Biotechnology, 18 (2):233-234. doi: 10.1038/72708 pmid: 10657137
[35]	Tisserat B, Galletta P, Jones D. 1990. Carpel Polymorphism in citrus fruit. Botanical Gazette, 151 (1):54-63. doi: 10.1086/337805 URL
[36]	Wang Bin, Liu Li-li, Wu Xiao-fang. 2019. Two misunderstandings of two kinds of separation ratio in Mendel's law of heredity. Bulletin of Biology, 54 (12):40-41. (in Chinese)
	王彬, 刘丽丽, 吴效芳. 2019. 孟德尔遗传定律中2种分离比的2个认识误区. 生物学通报, 54 (12):40-41.
[37]	Wang C Q, Zhao B L, He L L, Zhou S L, Liu Y, Zhao W Y, Guo S Q, Wang R R, Bai Q Z, Li Y H, Wang D F, Wu Q, Yang Y F, Liu Y, Tadege M, Chen J H. 2021. The WOX family transcriptional regulator SlLAM1 controls compound leaf and floral organ development in Solanum lycopersicum. Journal of Experimental Botany, 72 (5):1822-1835. doi: 10.1093/jxb/eraa574 URL
[38]	Wang Jing-yi, Chen Ye-yuan, Liu Wei-liang, Wu Yao-ting. 2008. Development and application of EST-SSR markers for bananas(Musa nana Lour). Hereditas, 30 (7):933-940. (in Chinese)
	王静毅, 陈业渊, 刘伟良, 武耀廷. 2008. 香蕉EST-SSRs标记的开发与应用. 遗传, 30 (7):933-940.
[39]	Wu H L, Chen D, Li J X, Yu B, Qiao X Y, Huang H L, He Y M. 2013. De novo characterization of leaf transcriptome using 454 sequencing and development of EST-SSR markers in tea (Camellia sinensis). Plant Molecular Biology Reporter, 31 (3):524-538. doi: 10.1007/s11105-012-0519-2 URL
[40]	Wu Han-yu. 2019. Study on function of the photoperiod response gene StYABBY1 in the regulation of photosynthesis[M. D. Dissertation]. Yinchuan: Ningxia University. (in Chinese)
	伍涵宇. 2019. 光周期响应基因StYABBY1调控光合作用功能研究[硕士论文]. 银川: 宁夏大学.
[41]	Yan Xiu-qin, Lu Min, An Hua-ming. 2015. Analysis on SSR information in transcriptome and development of molecular markers in Rosa roxburghii. Acta Horticulturae Sinica, 42 (2):341-349. (in Chinese)
	鄢秀芹, 鲁敏, 安华明. 2015. 刺梨转录组SSR信息分析及其分子标记开发. 园艺学报, 42 (2):341-349.
[42]	Yang Meng-ting, Huang Zhou, Gan Jian-ping, Xu Jun-chi, Pang Ji-liang. 2019. Research progress of SSR molecular markers. Journal of Hangzhou Normal University(Natural Science Edition), 18 (4):429-436. (in Chinese)
	杨梦婷, 黄洲, 干建平, 徐君驰, 庞基良. 2019. SSR分子标记的研究进展. 杭州师范大学学报(自然科学版), 18 (4):429-436.
[43]	You F M, Huo N X, Gu Y Q, Luo M C, Ma Y Q, Hane D, Lazo G R, Dvorak J, Anderson O D. 2008. Batchprimer3:a high throughput web application for PCR and sequencing primer design. BMC Bioinformatics, 9:253. doi: 10.1186/1471-2105-9-253
[44]	Yue X Y, Liu G Q, Zong Y, Teng Y W, Cai D Y. 2014. Development of genic SSR markers from transcriptome sequencing of pear buds. Journal of Zhejiang University Science B, 15 (4):303-312. doi: 10.1631/jzus.B1300240 URL
[45]	Zhang Qing-tian, Li Xiao-yan, Yang Yi-ming, Fan Shu-tian, Ai Jun. 2016. Analysis on SSR information in transcriptome and development of molecular markers in Lonicera caerulea. Acta Horticulturae Sinica, 43 (3):557-563. (in Chinese) doi: 10.16420/j.issn.0513-353x.2015-0659
	张庆田, 李晓艳, 杨义明, 范书田, 艾军. 2016. 蓝靛果忍冬转录组SSR信息分析及其分子标记开发. 园艺学报, 43 (3):557-563. doi: 10.16420/j.issn.0513-353x.2015-0659
[46]	Zhang Tian, Li Zuo-zhou, Liu Ya-ling, Jiang Zheng-wang, Huang Hong-wen. 2007. Genetic diversity gene introgression and homoplasy in sympatric populations of the genus Actinidia as revealed by chloroplast microsatellite markers. Biodiversity Science, 15 (1):1-22. (in Chinese) doi: 10.1360/biodiv.060277 URL
	张田, 李作洲, 刘亚令, 姜正旺, 黄宏文. 2007. 猕猴桃属植物的cpSSR遗传多样性及其同域分布物种的杂交渐渗与同塑. 生物多样性, 15 (1):1-22. doi: 10.1360/biodiv.060277
[47]	Zhao Yujie, Liu Cuiyu, Zhao Xueqing, Wang Yuying, Yan Ming, Yuan Zhaohe. 2021. Cloning and spatiotemporal expression analysis of PgWUS and PgBEL1 in Punica granatum. Acta Horticulturae Sinica, 48 (2):355-366. (in Chinese) doi: 10.16420/j.issn.0513-353x.2020-0377
	赵玉洁, 刘翠玉, 招雪晴, 汪钰莹, 闫明, 苑兆和. 2021. 石榴花器官发育相关基因PgWUS和PgBEL1克隆及其时空表达分析. 园艺学报, 48 (2):355-366. doi: 10.16420/j.issn.0513-353x.2020-0377
[48]	Zong Yu, Wang Yue, Zhu You-yin, Shao Xu, Li Yong-qiang, Guo Wei-dong. 2016. Development and validation of SSR markers based on transcriptomic data of Chinese cherry (Prunus pseudocerasus). Acta Horticulturae Sinica, 43 (8):1566-1576. (in Chinese)
	宗宇, 王月, 朱友银, 邵姁, 李永强, 郭卫东. 2016. 基于中国樱桃转录组的SSR分子标记开发与鉴定. 园艺学报, 43 (8):1566-1576. doi: 10.16420/j.issn.0513-353x.2016-0005

引物序号 Primer code	重复单元 Repeat motif	引物序列（5′-3′） Primer sequence		产物大小/bp Product size
Z1	(AT)*8	F：<M13>-TAACCTAGCATCCTCCAC	R：TATGCGGGTAAATCTCGT	184
Z5	(TC)*10	F：<M13>-CATTGATTCTTTCCTTCG	R：AAAATTAGCCAGAAACGT	247
Z7	(TA)*9	F：<M13>-TGATCTCCTCTGGCGAATG	R：CCTCCTCGTCACCATCCA	277
Z15	(TC)*6	F：<M13>-TCCCCTTCATTTATCCTT	R：TTTTGTTGATGGCGTTGG	172
Z17	(CT)*8	F：<M13>-ATGCAAGCCGCTTTTCTC	R：ACCCAAGATTCTGGTCAGG	203
Z24	(AT)*18	F：<M13>-ACCAGTCCTACGCATCAC	R：CCCGATACCCTGGATTTT	260
Z52	(TA)*12	F：<M13>-CTCCCACTGTCCCTTCGT	R：GCTTGCTGAGCGGATTTA	186
Z54	(AT)*14	F：<M13>-CATTAGAGTTCATTGGGAATA	R：TACCTTACGTTTTGTCGC	254
Z92	(CT)*12	F：<M13>-TACGGTATCAATTCCTTC	R：GGCATTATCAGACCCAAA	312
Z124	(AG)*23	F：<M13>-CCGATCATCAGGGACTACTA	R：CATCTCCAGCACCATTCTT	316
Z131	(CT)*9	F：<M13>-TATAAGGCAAGTGGGGTGA	R：GTGGCAACAAGAATACAAGAT	161
Z171	(TA)*17	F：<M13>-CTAAAGCGATCTTGACATA	R：GGACGGGACTTACTACCAG	252
Z178	(AT)*11	F：<M13>-GAGAACTTTTGTTTATTGGAG	R：TTCTTTCATGCTTCCGTTC	165
Z198	(TAA)*13	F：<M13>-GTAACTGGTGGATTTGTCG	R：CCATAGTCGGACCCTCTTT	201
Z199	(TAA)*13	F：<M13>-GTAACTGGTGGATTTGTCG	R：CCATAGTCGGACCCTCTTT	220
Z204	(TG)*9	F：<M13>-TGCTGGAAAGGAACGAAAC	R：CATTGCATGGCTGCTCATA	244
M13通用引物 <FAM>或<HEX>-TGTAAAACGACGGCCAGT
M13 universal primer

引物序号 Primer code	重复单元 Repeat motif	引物序列（5′-3′） Primer sequence		产物大小/bp Product size
Z1	(AT)*8	F：<M13>-TAACCTAGCATCCTCCAC	R：TATGCGGGTAAATCTCGT	184
Z5	(TC)*10	F：<M13>-CATTGATTCTTTCCTTCG	R：AAAATTAGCCAGAAACGT	247
Z7	(TA)*9	F：<M13>-TGATCTCCTCTGGCGAATG	R：CCTCCTCGTCACCATCCA	277
Z15	(TC)*6	F：<M13>-TCCCCTTCATTTATCCTT	R：TTTTGTTGATGGCGTTGG	172
Z17	(CT)*8	F：<M13>-ATGCAAGCCGCTTTTCTC	R：ACCCAAGATTCTGGTCAGG	203
Z24	(AT)*18	F：<M13>-ACCAGTCCTACGCATCAC	R：CCCGATACCCTGGATTTT	260
Z52	(TA)*12	F：<M13>-CTCCCACTGTCCCTTCGT	R：GCTTGCTGAGCGGATTTA	186
Z54	(AT)*14	F：<M13>-CATTAGAGTTCATTGGGAATA	R：TACCTTACGTTTTGTCGC	254
Z92	(CT)*12	F：<M13>-TACGGTATCAATTCCTTC	R：GGCATTATCAGACCCAAA	312
Z124	(AG)*23	F：<M13>-CCGATCATCAGGGACTACTA	R：CATCTCCAGCACCATTCTT	316
Z131	(CT)*9	F：<M13>-TATAAGGCAAGTGGGGTGA	R：GTGGCAACAAGAATACAAGAT	161
Z171	(TA)*17	F：<M13>-CTAAAGCGATCTTGACATA	R：GGACGGGACTTACTACCAG	252
Z178	(AT)*11	F：<M13>-GAGAACTTTTGTTTATTGGAG	R：TTCTTTCATGCTTCCGTTC	165
Z198	(TAA)*13	F：<M13>-GTAACTGGTGGATTTGTCG	R：CCATAGTCGGACCCTCTTT	201
Z199	(TAA)*13	F：<M13>-GTAACTGGTGGATTTGTCG	R：CCATAGTCGGACCCTCTTT	220
Z204	(TG)*9	F：<M13>-TGCTGGAAAGGAACGAAAC	R：CATTGCATGGCTGCTCATA	244
M13通用引物 <FAM>或<HEX>-TGTAAAACGACGGCCAGT
M13 universal primer

基因 Gene	序列号 ID	引物序列（5′-3′） Primer sequence		产物大小/bp Product size
SEP1	Cm158770.1	F：GCTTAGGCTTAGAGGAGTTG	R：GAATGAGCAGGTTGGTTG	244
SEP2	Cm240370.1	F：AGCGAACTCAGAGGAATC	R：ACCTAGTTGCTGCTCACC	263
SEP4	Cm146780.1	F：CCTGAGGAAGAAGTTGGAC	R：CGATAGAGGCTGAAAGAACC	142
SHP1	Cm078390.1	F：GGAATCGGCAGAGTCAGA	R：GCGTACTGGTGATTGGGT	260
SHP2	Cm164940.1	F：AATACTGGGTCTATCTGTGAA	R：TCTCGGCTATCTTTGCTC	289
SPL6	Cm158790.1	F：GACTCATCTGTTGACCCTT	R：CACTACCGACTTCACCATTA	211
SPL8	Cm162720.1	F：AGATGAAGCGACTCAAGAAT	R：GTGGGTGGGAGCGTAAGA	286
SPL9	Cm119150.1	F：TCTGATCGACTTCAGTGCATATC	R：AATTCCAGGACCAGAGAAACC	208
YABBY1A	Cm015380.1	F：ATCCCAAATCCATCTCCGAAC	R：TGAATCGGTTGTACGCAGAG	157
YABBY1B	Cm112520.1	F：ACTTCCCTCACATCCATTTCG	R：ACTACAACGAAACCCCAGC	192
YABBY4	Cm075530.1	F：CAAACATGGCTCACAAGGAAG	R：CTTCAAAGTTCCAAGCTCGC	132
YABBY5A	Cm147040.1	F：TGCGAACACCCACTAACAAG	R：AATGGATGTGAGGGAAGTGTG	198
CRC	Cm158810.1	F：CTCTCACCACTTCTCTCTTCTTC	R：AGAAGTTGCAGCGGACATAG	204
WOX1	Cm275910.1	F：ATTCTACTACGACTGCTATT	R：TCTCTCTTGTTTTGTTCACC	190
PRS	Cm223590.1	F：CCATAAAGCAAGAGAGAGGC	R：AGCTGGGTTAGAAAAGGAAC	190
LUG	Cm111030.1	F：CCCACCTCCTAAATGGCAATA	R：TAGCCGCATCATCCAAAGAA	150
AGL66	Cm105600.1	F：ATGTGAAGGAGAAATGCTGG	R：AATCTTTGGGTTGAGACTGG	388
AP2	Cm077350.1	F：AATCTAAAATAATTGCCCAA	R：AATAACTCAACTCACTCTGC	324
TIR1	Cm233920.1	F：GACGCAGCTCTACTCTCAGG	R：CTTTCTTGGTCCAGCAACAG	204
CsActin	Cs1g05000.1	F：CCAAGCAGCATGAAGATCAA	R：ATCTGCTGGAAGGTGCTGAG	101

基因 Gene	序列号 ID	引物序列（5′-3′） Primer sequence		产物大小/bp Product size
SEP1	Cm158770.1	F：GCTTAGGCTTAGAGGAGTTG	R：GAATGAGCAGGTTGGTTG	244
SEP2	Cm240370.1	F：AGCGAACTCAGAGGAATC	R：ACCTAGTTGCTGCTCACC	263
SEP4	Cm146780.1	F：CCTGAGGAAGAAGTTGGAC	R：CGATAGAGGCTGAAAGAACC	142
SHP1	Cm078390.1	F：GGAATCGGCAGAGTCAGA	R：GCGTACTGGTGATTGGGT	260
SHP2	Cm164940.1	F：AATACTGGGTCTATCTGTGAA	R：TCTCGGCTATCTTTGCTC	289
SPL6	Cm158790.1	F：GACTCATCTGTTGACCCTT	R：CACTACCGACTTCACCATTA	211
SPL8	Cm162720.1	F：AGATGAAGCGACTCAAGAAT	R：GTGGGTGGGAGCGTAAGA	286
SPL9	Cm119150.1	F：TCTGATCGACTTCAGTGCATATC	R：AATTCCAGGACCAGAGAAACC	208
YABBY1A	Cm015380.1	F：ATCCCAAATCCATCTCCGAAC	R：TGAATCGGTTGTACGCAGAG	157
YABBY1B	Cm112520.1	F：ACTTCCCTCACATCCATTTCG	R：ACTACAACGAAACCCCAGC	192
YABBY4	Cm075530.1	F：CAAACATGGCTCACAAGGAAG	R：CTTCAAAGTTCCAAGCTCGC	132
YABBY5A	Cm147040.1	F：TGCGAACACCCACTAACAAG	R：AATGGATGTGAGGGAAGTGTG	198
CRC	Cm158810.1	F：CTCTCACCACTTCTCTCTTCTTC	R：AGAAGTTGCAGCGGACATAG	204
WOX1	Cm275910.1	F：ATTCTACTACGACTGCTATT	R：TCTCTCTTGTTTTGTTCACC	190
PRS	Cm223590.1	F：CCATAAAGCAAGAGAGAGGC	R：AGCTGGGTTAGAAAAGGAAC	190
LUG	Cm111030.1	F：CCCACCTCCTAAATGGCAATA	R：TAGCCGCATCATCCAAAGAA	150
AGL66	Cm105600.1	F：ATGTGAAGGAGAAATGCTGG	R：AATCTTTGGGTTGAGACTGG	388
AP2	Cm077350.1	F：AATCTAAAATAATTGCCCAA	R：AATAACTCAACTCACTCTGC	324
TIR1	Cm233920.1	F：GACGCAGCTCTACTCTCAGG	R：CTTTCTTGGTCCAGCAACAG	204
CsActin	Cs1g05000.1	F：CCAAGCAGCATGAAGATCAA	R：ATCTGCTGGAAGGTGCTGAG	101

重复类型	重复次数 Number of repeat									总数	比例/%
Repeat type	5	6	7	8	9	10	11	12	>12	Total	Percentage
二核苷酸Dinucleotide	—	712	415	283	243	166	101	59	150	2 129	43.37
三核苷酸Trinucleotide	1 322	605	311	131	49	35	18	8	22	2 501	50.95
四核苷酸Tetranucleotide	115	36	7	6	2	0	0	0	0	166	3.38
五核苷酸Pentanucleotide	20	8	3	0	0	0	0	0	0	31	0.63
六核苷酸Hexanucleotide	59	13	7	0	1	2	0	0	0	82	1.67
总数Total	1 516	1 374	743	420	295	203	119	67	172	4 909
比例/% Percentage	30.88	27.99	15.14	8.56	6.01	4.14	2.42	1.36	3.50		100.00

Identification of F₁ Hybrids and Analysis of Fruit Shape Difference Between Fingered Citron and Citron

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 13

References 48

Related Articles 15

Recommended Articles

Metrics

Comments

重复类型	重复基序	所占比例/%
Repeat type	Repeat motif	Percentage
二核苷酸	AG/CT	55.61
Dinucleotide	AC/GT	15.45
	AT/AT	21.18
	CG/CG	0.75
三核苷酸	AAG/CTT	24.32
Trinucleotide	AGG/CCT	4.37
	ACC/GGT	5.19
	AGC/CTG	11.69
	ATC/ATG	16.06
	CCG/CGG	3.30
	ACG/CGT	2.48
	AAC/GTT	7.20
	AAT/ATT	24.20
	ACT/AGT	1.18

引物序号 Primer code	N_a观测等位基因数 Observed number of alleles	N_e有效等位基因数 Effective number of alleles	H_o观测杂合度 Observed heterozygosity	H_e期望杂合度 Expected heterozygosity	P_i多态性信息量 Polymorphism information content
Z1	28	21	1.000	0.981	0.956
Z5	18	16	0.188	0.966	0.932
Z7	17	14	0.429	0.963	0.924
Z15	20	21	0.667	0.958	0.932
Z17	28	17	0.941	0.988	0.957
Z24	25	18	0.611	0.968	0.938
Z52	19	14	0.929	0.968	0.930
Z54	27	20	0.750	0.981	0.954
Z92	25	19	0.526	0.977	0.949
Z124	17	16	0.438	0.948	0.912
Z131	32	21	0.952	0.986	0.961
Z171	25	19	0.579	0.974	0.946
Z178	26	19	0.474	0.982	0.954
Z198	28	21	1.000	0.978	0.953
Z199	30	21	0.762	0.985	0.960
Z204	22	20	0.550	0.967	0.940
均值 Mean	24.188	18.563	0.675	0.973	0.944

基因 Gene	相关系数 Correlation coefficient		基因 Gene	相关系数 Correlation coefficient
基因 Gene	现瓣期 Petals emerging stage	即将开放期 Pre-anthesis stage	基因 Gene	现瓣期 Petals emerging stage	即将开放期 Pre-anthesis stage
SEP1	-0.358	0.086	YABBY4	0.501	0.258
SEP2	-0.645	-0.775^*	YABBY5A	0.645	0.430
SEP4	0.788^*	-0.430	CRC	0.358	0.775
SHP1	-0.072	-0.775^*	WOX1	0.788^*	-0.775^*
SHP2	-0.645	-0.775^*	PRS	0.620	0.430
SPL6	-0.358	0.775^*	LUG	-0.072	-0.775^*
SPL8	-0.501	-0.430	AGL66	-0.358	0.086
SPL9	0.501	0.258	AP2	-0.501	-0.602
YABBY1A	0.358	0.258	TIR1	0.501	0.775^*
YABBY1B	0.501	0.258

[1]	NIE Xinghua, ZHANG Yu, LIU Song, YANG Jiabin, HAO Yaqiong, LIU Yang, QIN Ling, and XING Yu. Study on Genetic Characteristics and Taxonomic Status of Wild Chinese Chestnut Based on Genome Re-sequencing [J]. Acta Horticulturae Sinica, 2023, 50(8): 1622-1636.
[2]	WANG Ping, SHENG Ling, YANG Jinpeng, ZHOU Linglei, JIN Yan, LUO Xuzhao, MA Xianfeng, DENG Ziniu. Evaluation of Resistance to Citrus Canker Disease in Hybrid Progeny of Red Pomelo and American Citron [J]. Acta Horticulturae Sinica, 2023, 50(4): 765-777.
[3]	SUN Zeshuo, JIANG Dongyue, LIU Xinhong, SHEN Xin, LI Yingang, QU Yufei, LI Yonghua. Cluster Analysis and Construction of DNA Fingerprinting of 42 Oriental Cultivars of Flowering Cherry Based on SSR Markers [J]. Acta Horticulturae Sinica, 2023, 50(3): 657-668.
[4]	WANG Rui, HONG Wenjuan, LUO Hua, ZHAO Lina, CHEN Ying, WANG Jun. Construction of SSR Fingerprints of Pomegranate Cultivars and Male Parent Identification of Hybrids [J]. Acta Horticulturae Sinica, 2023, 50(2): 265-278.
[5]	LIU Yiping, NI Menghui, WU Fangfang, LIU Hongli, HE Dan, KONG Dezheng. Association Analysis of Organ Traits with SSR Markers in Lotus（Nelumbo nucifera） [J]. Acta Horticulturae Sinica, 2023, 50(1): 103-115.
[6]	ZHANG Kai, MA Mingying, WANG Ping, LI Yi, JIN Yan, SHENG Ling, DENG Ziniu, MA Xianfeng. Identification of HSP20 Family Genes in Citrus and Their Expression in Pathogen Infection Responses Citrus Canker [J]. Acta Horticulturae Sinica, 2022, 49(6): 1213-1232.
[7]	MA Mingying, HAO Chenxing, ZHANG Kai, XIAO Guihua, SU Hanying, WEN Kang, DENG Ziniu, MA Xianfeng. CsSWEET2a Promotes the Infection of Xanthomonas citri subsp. citri [J]. Acta Horticulturae Sinica, 2022, 49(6): 1247-1260.
[8]	LI Chao, YANG Ying, CHEN Wei, ZHENG Heyun, LIAO Xinfu, SUN Yuping. Construction of DNA Fingerprinting and Clustering Analysis with SSR Markers for the Muskmelon of Xizhoumi Series [J]. Acta Horticulturae Sinica, 2022, 49(3): 622-632.
[9]	NIE Xinghua, LI Yiran, TIAN Shoule, WANG Xuefeng, SU Shuchai, CAO Qingqin, XING Yu, QIN Ling. Construction of DNA Fingerprint Map and Analysis of Genetic Diversity for Chinese Chestnut Cultivars（Lines） [J]. Acta Horticulturae Sinica, 2022, 49(11): 2313-2324.
[10]	CHEN Mingkun, CHEN Lu, SUN Weihong, MA Shanhu, LAN Siren, PENG Donghui, LIU Zhongjian, AI Ye. Genetic Diversity Analysis and Core Collection of Cymbidium ensifolium Germplasm Resources [J]. Acta Horticulturae Sinica, 2022, 49(1): 175-186.
[11]	SONG Yun, JIA Mengjun, CAO Yaping, LI Zheng, HE Jiaxin, WANG Yongfei, ZHANG Xinrui, QIAO Yonggang. Analysis on Chloroplast Genomic Characteristics of Forsythia suspensa [J]. Acta Horticulturae Sinica, 2022, 49(1): 187-199.
[12]	WANG Xin, LI Mingyang, TIAN Lin, LIU Dongyun. ISSR and rDNA-ITS Sequence Analysis of the Genetic Relationship of Clematis in Hebei Province [J]. Acta Horticulturae Sinica, 2021, 48(9): 1755-1767.
[13]	WANG Xiaoqing, MEI Yulin, LIU Jinlian, XU Jinjian, LI Yongqiang, CHEN Wenrong, ZONG Yu, SHAO Wenke, LIAO Fanglei, GUO Weidong. A New Fingered Citron Cultivar‘Cuizhi’ [J]. Acta Horticulturae Sinica, 2021, 48(8): 1639-1640.
[14]	DING Yunhua, Budahn Holger, ZHAO Hong, ZHAO Xiuyun. Accurate Identification of Radish Chromosome F in the Backcross Progeny of Brassica rapa and Rape-radish Chromosome F Addition Line [J]. Acta Horticulturae Sinica, 2021, 48(7): 1295-1303.
[15]	ZHAO Qing, DU Zhenzhen, LI Xixiang, SONG Jiangping, ZHANG Xiaohui, YANG Wenlong, JIA Huixia, WANG Haiping. Genetic Diversity of Garlic Germplasm Resources Based on SSRseq Molecular Markers [J]. Acta Horticulturae Sinica, 2021, 48(7): 1397-1408.