摘要
【目的】黄瓜是世界十大蔬菜之一,单性结实是与黄瓜产量和品质密切相关的重要性状。对黄瓜单性结实进行研究,探明全雌性单性结实性状的遗传规律,并进行QTL定位,为进一步了解其分子机理和标记辅助育种奠定基础,也为黄瓜单性结实性状改良提供理论依据。【方法】试验采用对单株主、侧蔓各夹8朵雌花,待所有单株夹花结束后8—10 d一次性调查单性结实座瓜的方法,通过计算单性结实百分率(单性结实瓜数/所夹雌花数×100%)来评价单性结实能力。利用全雌单性结实材料EC1和雌雄同株非单性结实材料8419、14519分别构建的F2群体进行遗传分析。以EC1×8419杂交得到的部分F2为作图群体,利用9930和gy14黄瓜全基因组测序开发的1 335对SSR标记和双亲重测序开发的143对Indel标记引物进行多态引物筛选,采用Jion Map4.0软件进行遗传图谱构建,以该配组F2﹕3家系群体为研究对象,利用Win QTLcart2.5软件进行单性结实QTL检测。运用生物信息学的分析方法对初定位主效QTL区间进行候选基因分析。【结果】试材EC1的单性结实遗传符合数量性状的特征,但与不同材料配组的后代群体分离偏向不同亲本。构建了一张含有7条染色体、116个SSR标记和9个Indel标记的连锁图谱,图谱总长度为802.9 c M,标记间平均距离6.3 c M。共检测到7个与单性结实相关的QTL,分别为Parth1、Parth2-1、Parth2-2、Parth3-1、Parth3-2、Parth5、Parth7,分布在1、2、3、5、7染色体上。其中仅Parth2-1在春、秋两季中均被检测到,LOD值分别为9.0和6.2,贡献率为17.4%和10.2%,位于标记SSR00684-SSR22083之间,认为是控制单性结实的主效QTL位点。该区段的遗传距离为17.1 c M,物理距离2.9 Mb,包含307个基因,推测参与植物激素信号转导的基因Csa2M035330.1和Csa2M070880.1是与单性结实相关性较大的候选基因。其他位点都为微效位点。【结论】单性结实的遗传符合数量性状特征,位于2号染色体上的Parth2-1是控制黄瓜单性结实性状的主效QTL位点。推测植物激素代谢通路中的基因是可能的候选基因。研究结果为单性结实主效基因的精细定位和克隆及分子标记辅助育种奠定了基础。
【Objective】Cucumber is one of the ten vegetables in the world and parhenocarpy is an important trait closely related to production and quality of cucumber.To explore the inheritance and QTL mapping for parthenocarpy in cucumber could provide a preliminary basis for further study on mechanism of parthenocarpy and molecular assistant selection breeding,and lay a theoretical foundation for breeding of parthenocarpy.【 Method 】 In this study,The authers clipped eight female flowers on main stem and branches,respectively,for every individual plant and investigated parthenocarpic fruit once when all plants treament finished 8-10 days later to calculate the parthenocarpy percentage(numbers of parthenocarpis fruit/numbers of clipped female flower) in order toevaluate parthenocarpy ability.Two F2 progenies derived from two crosses between EC1,a gynoecious parthenocarpic line,and two monoecious non-parthenocarpic lines 8419 and 14519 were constructed to determine the inheritance of parthenocarpy in cucumber.A linkage map from part of F2 plants from the cross of EC1×8419 was constructed with Join Map4.0 software by screening 1 335 SSR from 9930 and gy14 cucumber genome sequencing and 143 Indel primers from two parents resequencing and QTL detection for parthenocarpy was conducted with Win QTLcart2.5 software using F2:3 families from the same cross.The candidate genes in major QTL region were predicted using bioinformatic analysis method.【 Result 】 Parthenocarpy in EC1 was inherited quantitatively but segregated towards different parents in two F2 progenies.A linkage map containing 7 chromosomes,116 SSR and 9 Indel markers was constructed,which total length was 802.9 c M and average distance between two markers was 6.3 c M.QTL analysis identified 7QTLs,Parth1,Parth2-1,Parth2-2,Parth3-1,Parth3-2,Parth5,and Parth7,distributing on chromosomes 1,2,3,5,and 7.The major QTL Parth2-1 locating between SSR00684-SSR22083 was the only locus detected in two seasons,having two LOD scores of 9.0,6.2and R2 of 17.4%,10.2%,respectively,and its genetic and physical distance was 17.1 c M and 2.9 Mb.There were 307 genes in this region and two gene among them,Csa2M035330.1 and Csa2M070880.1,involved in plant hormone signal transduction would be candidate genes closely related to parthenocarpy,the rest were minor QTLs.【 Conclusion 】 The inheritance of pathenocarpy was quantitative.The QTL Parth2-1 locating on chromosome 2 was the major QTL controlling parthenocarpy in cucumber and two genes in plant hormone pathway would be candidate genes.The results in this study would lay a foundation for fine mapping and gene cloning of major QTL of parthenocarpy in cucumber and for use in MAS breeding.
引文
[1]陈学好,曹碚生.黄瓜单性结实研究概况.中国蔬菜,1994(3):56-59.Chen X H,Cao B S.Research on parthenocarpy in Cucumber.China Vegetables,1994(3):56-59.(in Chinese)
[2]Hawthorn L R,Wellington R.Geneva,a greenhouse cucumber that develops fruit without pollination.New York Agricultural Experiment Station Bulletin,1930,580:1-11.
[3]Juldasheva L M.Inheritance of the tendency towards parthenocarpy in cucumbers.Byull.vsesoyuznogo ordena Lenina Inst.Rastenievodstva Imeni N.I.Vavilova,1973,32:58-59.
[4]Meshcherov E T,Juldasheva L M.Parthenocarpy in cucumber.Trudy-po-Prikladnoi-Botanike-Genetiki-I-Selektsii,1974,51:204-213.
[5]Pike L M,Peterson C E.Inheritance of parthenocarpy in the cucumber(Cucumis sativus L.).Euphytica,1969,18(1):101-105.
[6]Ponti O M B,Garretsen F.Inheritance of parthenocarpy in pickling cucumbers(Cucumis sativus L.)and linkage with other characters.Euphytica,1976,25(1):633-642.
[7]El Shawaf I I S,Baker L R.Inheritance of parthenocarpic yield in gynoecious pickling cucumber for onec-over mechanical harvest by diallel analysis of six gynoecious lines.Journal of the American Society for Horticultural Science,1981,106(3):359-364.
[8]El Shawaf I I S,Baker L R.Combining ability and genetic variances of G×H F1 hybrids for parthenocarpic yield in gynoecious pickling cucumber from once-over mechanical harvest.Journal of the American Society for Horticultural Science,1981,106(3):365-370.
[9]曹碚生,陈学好,徐强,顾春山.黄瓜单性结实世代遗传效应的初步研究.园艺学报,1997,24(1):53-56.Cao B S,Chen X H,Xu Q,Gu C S.The genetic effects of parthenocarpic generations of cucumber.Acta Horticulturae Sinica,1997,24(1):53-56.(in Chinese)
[10]Sun Z Y,Lower R L,Staub J E.Analysis of generation means and components of variance for parthenocarpy in cucumber(Cucumissativus L.).Plant Breeding,2006,125(3):277-280.
[11]闫立英,娄丽娜,娄群峰,陈劲枫.全雌黄瓜单性结实性的遗传分析.园艺学报,2008,35(10):1441-1446.Yan L Y,Lou L N,Lou Q F,Chen J F.Inheritance of parthenocarpy in gynoecious cucumber(Cucumis sativus L.).Acta Horticulturae Sinica,2008,35(10):1441-1446.(in Chinese)
[12]闫立英,娄丽娜,李晓丽,娄群峰,冯志红,陈劲枫.雌雄同株黄瓜单性结实性遗传分析.中国农业科学,2010,43(6):1295-1301.Yan L Y,Lou L N,Li X L,Lou Q F,Feng Z H,Chen J F.Inheritance of parthenocarpy in monoecious cucumber(Cucumis sativus L.).Scientia Agricultura Sinica,2010,43(6):1295-1301.(in Chinese)
[13]王莉莉,司龙亭,邹芳斌.黄瓜单性结实的遗传分析.湖北农业科学,2008,47(4):437-439.Wang L L,Si L T,Zou F B.Genetic analysis of parthenocarpy incucumber.Hubei Agricultural Sciences,2008,47(4):437-439.(in Chinese)
[14]Sun Z Y,Staub J,Chung S M,E Lower R L.Identification and comparative analysis of quantitative trait loci associated with parthenocarpy in processing cucumber.Plant Breeding,2006,125(3):281-287.
[15]Cavagnaro P F,Senalik D A,Yang L M,Simon P W,Harkins T T,Kodira C D,Huang S W,Weng Y Q.Genome-wide characterization of simple sequence repeats in cucumber(Cucumis sativus L.).BMC Genomics,2010,11(1):1471-2164.
[16]Huang S W,Li R Q,Zhang Z H,Li L,Gu X F,Fan W,Lucas W J,Wang X W,Xie B Y,Ni P X,Ren Y,Zhu H M,Li J,Lin K,Jin W W,Fei Z J,Li G C,Staub J B,Kilian A,van der Vossen E A G,Wu Y,Guo J,He J,Jia Z Q,Ren Y,Tian G,Lu Y,Ruan J,Qian W,Wang M W,Huang Q F,Li B,Xuan Z L,Cao J J,San A,Wu Z G,Zhang J B,Cai Q L,Bai Y Q,Zhao B W,Han Y H,Li Y,Li X F,Wang S H,Shi Q X,Liu SQ,Cho W K,Kim J Y,Xu Y,Heller-Uszynska K,Miao H,Cheng Z C,Zhang S P,Wu J,Yang Y H,Kang H X,Li M,Liang H Q,Ren X L,Shi Z B,Wen M,Jian M,Yang H L,Zhang G J,Yang Z T,Chen R,Liu S F,Li J W,Ma L J,Liu H,Zhou Y,Zhao J,Fang X D,Li G Q,Fang L,Li Y G,Liu D Y,Zheng H K,Zhang Y,Qin N,Li Z,Yang G H,Yang S,Bolund L,Kristiansen K,Zheng H C,Li S C,Zhang X Q,Yang H M,Wang J,Sun R F,Zhang B X,Jiang S Z,Wang J,Du Y C,Li S G.The genome of the cucumber,Cucumis sativus L.Nature Genetics,2009,41(12):1275-1281.
[17]Zhang W W,Pan J S,He H L,Zhang C,Li Z,Zhao J L,Yuan X J,Zhu L H,Huang S W,Cai Run.Construction of a high density integrated genetic map for cucumber(Cucumis sativus L.).Theoretical and Applied Genetics,2012,124(2):249-259.
[18]Li H,Hearne S,Banziger M,Li Z,Wang J.Statistical properties of QTL linkage mapping in biparental genetic populations.Heredity,2010,105(3):257-267.
[19]李慧慧,张鲁燕,王健康.数量性状基因定位研究中若干常见问题的分析与解答.作物学报,2010,36(6):918-931.Li H H,Zhang L Y,Wang J K.Analysis and answers to frequently asked questions in quantitative trait locus mapping.Acta Agronomica Sinica,2010,36(6):918-931.
[20]Gustafson F G.Auxin distribution in fruits and its significance in fruit development.American Journal of Botany,1939,26(4):189-194.
[21]Beyer E M,Quebedeaux B.Parthenocarpy in cucumber:mechanism of action of auxin transport inhibitors.Journal of the American Society for Horticultural Science,1974,99:385-390.
[22]Mapelli S,Frova C,Torti G,Soressi G P.Relationship between set,development and activities of growth regulators in tomato fruits.Plant Cell Physiology,1978,19(7):1281-1288.
[23]Gillaspy G,Ben-David H,Gruissem W.Fruits:a developmental perspective.Plant Cell,1993,5(10):1439-1451.
[24]Vivian-Smith A,Koltunow A M.Genetic analysis of growth regulator-induced parthenocarpy in arabidopsis.Plant Physiology,1999,121(2):437-451.
[25]Serrani J C,Fos M,Atarés A,García-Martínez J L.Effect of gibberellin and auxin on parthenocarpic fruit growth induction in the cv.Micro-Tom of tomato.Journal of Plant Growth Regulation,2007,26(3):211-221.
[26]Fu F Q,Mao W H,Shi K,Zhou Y H,Asami,Yu J Q.A role of brassinosteroids in early fruit development in cucumber.Journal of Experimental Botany,2008,59(9):2299-2308.
[27]De-Jong M,Mariani C,Vriezen W H.The role of auxin and gibberellin in tomato fruit set.Journal of Experimental Botany,2009,60(5):1523-1532.