73份草莓种质资源表型性状的遗传多样性分析及在湖北省的综合评价.pdf

DOI:10.13925/ki.gsxb.2022067373份草莓种质资源表型性状的遗传多样性分析及在湖北省的综合评价肖桂林1，2，管自豪3，张思思4，温昕5，陈锡洋1，2，曾祥国1，2，张庆华1，2，刘爽1，5，刘思佳1，5，王涌1，2，邓江丽1，2，韩永超1，2*（1湖北省农业科学院经济作物研究所，武汉 430064；2蔬菜种质创新与遗传改良湖北省重点实验室，武汉 430064；3长江大学园艺园林学院，湖北荆州 434000；4上饶师范学院，江西上饶 334001；5华中农业大学，武汉 430070）摘要：【目的】全面系统地鉴定草莓种质资源表型变异，明确表型性状的遗传多样性，并基于综合评价指标筛选出优异资源，为草莓品种改良及理论研究奠定材料基础。【方法】对来源于国内、日韩及欧美的73份草莓种质资源的58个植物学、产量及品质性状进行系统评价和分析。【结果】51个表型性状具有不同程度的多态性，包括34个描述型性状和17个数值型性状。描述型性状共检测出115个变异类型，平均遗传多样性指数H 达到0.85；数量型性状遗传多样性指数H 的变异范围为1.682.92，表明供试草莓资源性状变异丰富，尤其是果实性状，平均遗传多样性指数（1.05）远高于非果实性状（0.61）。PCA分析结果均表明，日韩草莓与中国草莓在表型上较为相似，欧美草莓与亚洲草莓在表型上存在较大差异。进一步通过综合指标评价值D鉴定出7份优等种质，长势和产量均强于和高于其他种质。【结论】初步构建了草莓种质资源在湖北省的综合评价体系，为草莓品种的遗传改良奠定了基础。关键词：草莓；种质资源；表型性状；遗传多样性中图分类号：S668.1文献标志码：A文章编号：1009-9980(2023)08-1546-13收稿日期：2022-12-04接受日期：2023-03-22基金项目：国家重点研发计划（2019YFD1000803）；湖北省重点研发计划（2021BBA099，2022BBA085）；湖北省农业科技创新中心资助项目（2021-620-000-001-008）作者简介：肖桂林，女，助理研究员，研究方向为草莓分子育种。Tel：[phone]，E-mail：*通信作者Author for correspondence.Tel：[phone]，E-mail：果树学报2023，40（8）：1546-1558Journal of Fruit ScienceGenetic diversity and comprehensive evaluation of phenotypic traits in73 germplasm resources of cultivated strawberries grown in Hubei prov-inceXIAO Guilin1,2,GUAN Zihao3,ZHANG Sisi4,WEN Xin5,CHEN Xiyang1,2,ZENG Xiangguo1,2,ZHANG Qinghua1,2,LIU Shuang1,5,LIU Sijia1,5,WANG Yong1,2,DENG Jiangli1,2,HAN Yongchao1,2*(1Institute of Industrial Crops,Hubei Academy of Agricultural Sciences,Wuhan 430064,Hubei,China;2Hubei Key Laboratory of Vegeta-ble Germplasm Enhancement and Genetic Improvement,Wuhan 430064,Hubei,China;3College of Horticulture and Gardening,YangtzeUniversity,Jingzhou 434000,Hubei,China;4Shangrao Normal University,Shangrao 334001,Jiangxi,China;5Huazhong AgriculturalUniversity,Wuhan 430070,Hubei,China)Abstract:【Objective】The purpose of this study is to explore the genetic diversity of cultivated straw-berry germplasms from China,Japan and Korea(JapKor),Europe and America(Europe)and identifyexcellent germplasms by comprehensively screening phenotypic traits,including morphological traits,yield,and fruit quality,which will lay the foundation for the molecular improvement and genetic map-ping of important agronomic traits in cultivated strawberry.【Methods】Based on strawberry germplasmresource descriptors,58 phenotypic traits were systematically investigated,including plant height,plantarchitecture,leaf traits,flower traits,runner,fruit and other related traits.Then,the coefficient of varia-tion as well as the Shannons diversity index(H)were calculated for each trait to estimate genetic di-versity.Next,the Pearson correlation analysis,the principal component analysis as well as the compre-，等：分析及在湖北省的综合评价第8期hensive evaluation value D were calculated by using the SPSS,SIMCA and R software.【Results】Among all 58 phenotypic traits,except for petiole color(olivine),calyx color(green),blade margin ser-rated(sharp),lobular number(three),flower sex(hermaphrodite flower),petal shape(round),degree ofconcavity of calyx center(concave),the remaining 51 traits showed different degrees of phenotypic vari-ation,of which 34 traits were descriptive traits and 17 traits were quantitative traits.There were 115variation types for 34 descriptive traits,and 3.38 variation types for each descriptive trait.The Shannons diversity index H ranged from 0.24 to 1.49,with an average score of 0.85.The coefficient of varia-tion of the 17 quantitative traits ranged from 10.31%to 60.65%and the Shannon s diversity indexranged from 1.68 to 2.92.Genetic diversity analysis showed that strawberry germplasm resourcesshowed rich phenotypic variation.Furthermore,fruit traits showed more phenotypic variation than non-fruit traits because the average Shannon s diversity index H was 1.05 and 0.61 for fruit traits and non-fruit traits,respectively.The Shannons diversity index Hwas the highest for fruit flavor(1.49),fol-lowed by flesh color,whose Hwas 1.46.The third highest attribute was 1.35,which corresponded topulp texture and pulp color.The average fruit weight,fruit longitudinal diameter and fruit transverse di-ameter were 14.34 g,35.38 mm and 29.21 mm,respectively.The correlation coefficients among traitsranged from-0.70 to 0.95,the plant height and petiole length were extremely significantly and positive-ly correlated,with a correlation coefficient of 0.95.Petiole length was negatively correlated with plantarchitecture and the lowest correlation coefficient was-0.70.In addition,17 quantitative traits were pos-itively correlated with each other,i.e.,plant height,canopy diameter,leaf length,leaf width,length/width ratio,petiole length,petiole diameter,number of compound leaves,leaf thickness,pedicel length,pedicel diameter,flower numbers,flower diameter,plant architecture,fruit weight,fruit longitudinal di-ameter and fruit transverse diameter.Leaf length/width ratio was significantly correlated with leafshape,and trichome density of the upper leaf surface was significantly correlated with that of the lowerleaf surface and leaf texture,respectively.Principal component analysis transformed 51 phenotypictraits into 15 comprehensive factors,with a cumulative contribution rate of 78.14%.The contributionrates of the first three principal components were 15.92%,8.82%and 7.76%,respectively.The loadingsof the first principal components were plant height,canopy diameter,plant architecture and other traitsrepresenting growth vigor.The loadings of the second principal components featured fruit quality,likeflesh color and pulp texture.The loadings of the third principal components were flower numbers,fruitweight,fruit longitudinal diameter and fruit transverse diameter representing fruit yield.The results ofprincipal components also illustrated that strawberry resources of China and JapKor had very similarphenotypic traits and belonged to the Asian group,while the other germplasms belonged to the Europegroup,indicating that the germplasms had more phenotypic variations between the Asia group and Eu-rope group.Germplasm resources were divided into three categories according to the comprehensiveevaluation value D:excellent,medium and poor.A total of seven excellent germplasms were identified,4 of which were from China,namely Ganlu,Ningfeng,Miaoxiang 7 and 07-D-5.Three were from Jap-Kor including Kaorino,Benihoppe and TouKun.These excellent germplasms exhibited higher plantheight,bigger canopy diameter,heavier fruit weight,bigger fruit longitudinal and transverse diametersthan the other medium and poor germplasms.Among them,Kaorino showed the largest fruit longitu-dinal and transverse diameters and TouKun had the most flower numbers.In summary,these excellentstrawberry germplasms have good plant vigor and fruit yield,and have potential for breeding and theo-retical research.【Conclusion】A total of 73 cultivated strawberries grown in Hubei province showedabundant phenotypic diversity,especially for fruit traits.Strawberries from different geographical ori-肖桂林，等：73份草莓种质资源表型性状的遗传多样性分析及在湖北省的综合评价1547果树学报第40卷草莓（Fragaria ananassa Duch.）是蔷薇科草莓属多年生草本植物，果实鲜美红嫩、香气浓郁、酸甜可口，素有“水果皇后”的美誉。中国是世界第一大草莓生产国和消费国，2020年中国草莓种植面积12.7万hm2，产量达334万t（FAO，2020，http:/www.fao.org/statistics/en/），占世界种植面积和产量的1/3，年总产值超600亿元。然而，目前国内主栽草莓品种多为从国外引入，自主选育优良品种的缺乏是国内草莓产业发展中的瓶颈。种质资源是新品种选育、功能基因组学研究和生产实践的物质基础，全面系统地评价种质资源对遗传改良研究十分重要。表型作为最直观、快速的评价指标，指的是能够反映植物细胞、组织、器官、植株和群体的结构及功能特征的物理、生理和生化性质，其本质实际是植物基因图谱的时序三维表达及其地域分异特征和代际演进规律1。表型研究在植物系统研究中占据决定性地位，作为多组学研究中的重要组成成分之一，表型组学结合基因组学、转录组学，可以有效提速功能基因组学和分子育种研究2。草莓属植物倍性复杂，变异丰富，共包含24个种3-4，其中二倍体种12个、四倍体种5个、五倍体和六倍体种各 1 个、八倍体种 3 个以及十倍体种 2个5-6。现代栽培草莓是18世纪中期在法国由来自北美的八倍体弗吉尼亚草莓（F.virginiana）和来自南美的八倍体智利草莓（F.chiloensis）偶然杂交而成的异源八倍体7-9，遗传特性复杂，种质资源性状评价及遗传基础研究报道较少10，已有研究主要集中在抗病性11-13、抗逆性14-16及果实品质鉴定方面17-18，缺乏包括农艺性状在内的全面系统的鉴定及评价。笔者在本研究中以73份不同地理来源的栽培种草莓资源为材料，通过对58个表型性状进行鉴定，分析其遗传多样性并建立综合评价体系，研究结果将为草莓资源利用、品种选育及重要性状遗传改良研究奠定基础。1材料和方法1.1供试草莓材料供试材料为湖北省农业科学院经济作物研究所长期保存的73份栽培草莓资源（表1），其中37份来源于国内，16 份来源于日韩两国，23 份来源于欧美。每份资源材料定植20株用于性状鉴定，于2021年9月初定植于湖北省农业科学院经济作物研究所草莓基地双层塑料薄膜大棚内，起垄定植，垄高1520 cm，垄宽60 cm，一垄双行，株距20 cm。肥、水、温湿度及病虫害防治按常规方法管理。1.2表型性状鉴定表型性状鉴定严格按照 草莓种质资源描述规范19进行，于盛花期（11月初）分别选取5株测定各材料的株高、冠径、植株姿态、叶片长、叶片宽、叶片长宽比、叶柄长度、叶柄粗度、复叶数量、叶片厚、花梗长、花梗粗、花数、花冠径、雄蕊数、叶面状态、叶片颜色、叶片形状、叶片质地、托叶颜色、耳叶、花瓣数、花序高低、花序着生状态、花色、花瓣相对位置、雄蕊高低、叶正面茸毛密度、叶背面茸毛密度、叶柄茸毛密度、叶柄颜色、叶片边缘锯齿、小叶数、花性、花瓣形状共计35个性状，于12月初对果实性状进行调查，每份材料选取成熟度一致的果实5个，分别对果质量、果实纵径、果实横径、萼下着色、宿萼着生状态、宿萼颜色、萼心、无种子带、果形、果面状态、果面光泽、果面颜色、果尖着色、种子颜色、种子密度、种子着生状态、果肉颜色、髓心颜色、髓心大小、髓心空洞、香气、果肉质地、风味共计23个性状进行鉴定，其中41个描述型性状及赋值见表2。1.3数据统计与分析采用IBM SPSS统计各性状变异系数、均值及标准差等描述型指标及相关性分析，利用R 4.0.3 进行聚类分析及相关性热图绘制；利用 Excel 计算Shannon s diversity index（遗传多样性指数，H），计算公式为H=-Pi ln Pi，参考文献20-22的方法。利用SIMCA14.1进行主成分分析，利用隶属函数产生综合指标评价值D评价草莓种质资源23。隶属函数值：（Xi）=（XiXmin）/（XmaxXmin），i=1，2，n。（1）gins were clustered into different groups in principal component analysis based on phenotypic data.There may be more phenotypic variances between the Asia group and Europe group.Finally,seven ex-cellent strawberry germplasms were screened out through the comprehensive evaluation index D,whichcan be used as breeding parents and materials for theoretical research.Key words:Strawberry;Germplasm resources;Phenotypic traits;Genetic diversity1548，等：分析及在湖北省的综合评价第8期表 1供试的 73 份栽培种草莓种质资源及来源Table 1Seventy-three cultivated strawberry germplasm resources and origin来源Origin中国China日本Japan编号Number12345678910111213141516171819202122232425262728293031323334353637种质名称Name京桃香Jingtaoxiang九天红韵Jiutianhongyun京凝香Jingningxiang3公主Third Princess红玉Hongyu京郊小白Jingjiaoxiaobai京藏香Jingzangxiang晶硕Jingshuo天香Tianxiang京怡香Jingyixiang甘露Ganlu宁丰Ningfeng白雪公主Snow White黔莓1号Qianmei No.1粉红佳人Fenhongjiaren俏佳人Pretty Beauty宁玉Ningyu硕丽Shuoli白草莓Baicaomei晶玉Jingyu秀丽Xiuli越心Yuexin粉红公主Fenhonggongzhu粉佳人Pink Beauty晶瑶Jingyao京泉香Jingquanxiang宁玉-转基因Ningyu-cripsr-妙香7号Miaoxiang No.7丹莓2号Danmei No.2太空2008Taikong2008玉兔Yutu越秀Yuexiu晶悦Jingyue07-D-5艳丽Yanli佐贺清香Sagahonoka白雪小叮Baixuexiaoding来源Origin日本Japan韩国Korea美国America西班牙Spain加拿大Canada德国Germany荷兰Netherland法国France编号Number383940414243444546474849505152535455565758596061626364656667686970717273种质名称Name鬼努甘Kinuama丰香Toyonoka圣雪Shengxue章姬Akihime香野Kaorino红颜Benihoppe桃薰Toukun幸香Sachinoka枥乙女Tochiotome宝交早生Hokowase点雪Dianxue圣诞红Ssanta雪里香Xuelixiang特尼拉 Tenira蒙特瑞 Monterey圣安德瑞斯SanAndreas波特拉Portola卡姆罗莎Camorosa阿尔滨Albion哈尼Honeoye全明星Allstar派扎罗PajaroGento131CV111CV甜查理Sweet CharlieBF3美13 Honai吐德拉Tudla卡尔特Kart弗杰利亚Fujiliya威斯塔尔VeestarBURPEE森加.森加那Senga Sengana里瓦Riva达赛莱克特Darselect式中，Xi为第i个公因子的得分值，Xmin为第i个公因子得分的最小值，Xmax为第i个公因子得分的最大值。综 合 指 标 权 重：Wi=Pi/Pi，i=1，2，n。（2）式中，Wi为第i个公因子在所有公因子中的权重，Pi为各品种第i个公因子的贡献率。综合指标评价值：Dj=（Xi）Wi，j=1，2，k。（3）式中，k为样本数。2结果与分析2.1表型性状多样性评价2.1.1描述型性状共鉴定了73份草莓资源的58个表型性状，其中7个性状无变异，分别是叶柄颜色（黄绿）、叶片边缘锯齿（尖）、小叶数（3）、花性（两肖桂林，等：73份草莓种质资源表型性状的遗传多样性分析及在湖北省的综合评价1549果树学报第40卷表 2草莓 41 个描述型性状及赋值Table 2Forty-one descriptive traits and their assignment植株姿态Plant architecture（PA）叶面状态Foliar state（Foliar S）叶柄颜色Petiole color（Pet C）叶片颜色Leaf color（LC）叶片形状Leaf shape（Lshape）叶片质地Leaf texture（LText）托叶颜色Stipule color（Stipule C）耳叶Ear lobes（EL）花瓣数Petal numbers（PN）花序高低Inflorescences higher or lowerthan leaf（Inflo H L）花序着生状态InflorescencesArchitecture（InfloA）花色Flower color（Flower C）花瓣相对位置Relative position of petals（RPP）雄蕊高低Stamen higher or lower thanpistil（SHL）叶正面茸毛密度Trichome density of the upper leafsurface（TDULS）叶背面茸毛密度Trichome density of the lower leafsurface（TDLLS）叶柄茸毛密度Trichome density of petiole（TDP）萼下着色Subcalyx coloration（SubcalyxC）宿萼着生状态Persistent calyx（PC）无种子带Seedless belt（SB）果形Fruit shape（FS）果面状态Fruit surface state（FSS）果面光泽Fruit surface gloss（FSG）果面颜色Fruit surface color（FSC）果尖着色Fruit tip coloring（FTC）无None直立Erect匙状Key shape黄绿Olivine黄绿Olivine圆形Round柔软Soft浅绿Light Green平展Flat低于叶面Lower直立Erect白White相离Separate低于雌蕊Lower密Dense密Dense密Dense易Easy平贴Appressed无None圆锥形Conical平整Flat弱Weak白White易Easy中间Semi erect边向上Edge up紫红Magenta绿Green椭圆形Ellipse革质粗糙Leatheryand coarse浅红Pale red漏斗状Funnel-shaped平于叶面Same斜生Slanting粉红Pink相接Adjacent平于雌蕊Same中Medium中Medium中Medium中Medium平离Flat小Small长圆锥形Long conical浅沟少Less shallowgroove中Medium橙红Orange-red中Medium开张Patulous平Flat深绿Dark green菱形Rhombus革质平滑Leathery andsmooth深红Dark red兼有Both高于叶面Higher红Red重叠Overlapped高于雌蕊Higher疏Sparse疏Sparse疏Sparse难Difficult主萼平离副反卷Primary calyx flatfrom secondary tract中Medium短圆锥形Short conical浅沟多More Shallowgroove强Strong红Red难Difficult平而尖向下Flat and point down蓝绿Blue green卵圆形Oval反卷Rolling大Big楔形Wedge深沟少Less deep groove深红Dark red边向下Edge down倒卵圆形Obovate5畸形Misshapen深沟多More deepgroove紫红Magenta67性状 Trait各类型赋值Assignment of each number012345671550，等：分析及在湖北省的综合评价第8期种子颜色Seed color（SeedC）种子密度Seed density（SD）种子着生状态Seed-bearing condition（SBC）果肉颜色Flesh color（FleshC）髓心颜色Pith color（PithC）髓心大小Pith size（PiS）髓心空洞Hollow pith（HP）香气Fruit aroma（FA）果肉质地Pulp texture（PT）风味Fruit flavor（FF）宿萼颜色Calyx color（CalyxC）叶片边缘锯齿Blade margin serrated（BMS）小叶数Lobular number（LN）花性Flower sex（FSex）花瓣形状Petals shape（PS）萼心凹凸程度Degree of concavity of calyxcenter（CCCD）黄Yellow稀Sparse凹Concave白White白White小Small无None无None绵Mellowest酸Sour绿Green尖Sharp3Three两性hermaphroditeflower扁圆形Oblate凹Concave黄绿Olivine中Medium微凹Little concave橙黄Tiny orange橙黄Tiny orange中Medium小Small淡Faint松Spongy甜酸Sweet-sour枯黄Dry yellow钝Blunt5Five雌花Female flower圆形Round平Flat红Red密Dense平Flat橙红Orange-red橙红Orange-red大Big中Medium浓Thick韧Tougher酸甜适中Moderate sourand sweet3和5Three and five雄花Male flower扇形Fan-shaped凸Convex兼有Both微凸Little convex红Red红Red大Big脆Crisp酸甜Sour-sweet卵形Oval凸Convex深红Dark red深红Dark red甜Sweet纺锤形Spindly性状 Trait各类型赋值Assignment of each number01234567表 2（续）Table 2(Continued)性）、花瓣形状（圆）、宿萼颜色（绿）、萼心（凹），其余51个性状呈现不同程度的表型多态性，包含34个描述型性状和17个数值型性状（图1）。描述型性状包含16个植物学相关性状和18个果实性状，共计检测出34个性状的115个变异类型，平均每性状变异类型数为3.38个。其中，果形、果面状态、种子着生状态、果肉颜色、髓心颜色、风味变异类型数最多，有5个；叶片质地、花序高低和花序着生状态的变异类型数最少，各2个。遗传多样性指数H 的变异范围为0.241.49，均值达到0.85，表明供试草莓资源描述型性状变异丰富。果实性状的平均遗传多样性指数为1.05，其中果实风味的遗传多样性指数最高，为1.49；其次是果肉颜色，遗传多样性指数为1.46；果肉质地和髓心颜色的遗传多样性指数为1.35。遗传多样性指数最低的是无种子带，为0.62。与果实性状相比，非果实性状的遗传多样性指数显著低于果实性状，仅0.61。其中，植株姿态和叶正面茸毛密度遗传多样性指数最高，均为1.01，其次是叶片颜色和托叶颜色，为0.80。花色和叶柄茸毛密度的遗传多样性指数最低，为0.24，表明果实性状多样性更为丰富。2.1.2数值型性状17个数量性状变异系数范围为 10.31%60.65%（表 3），遗传多样性指数达到1.682.92，表明供试草莓资源数值型性状变异丰富。3个果实性状：果质量、果实纵径和横径中，单果质量的变异系数最大，为41.09%，平均单果质量14.34 g；果实纵径、横径变异系数相较于单果质量偏小，平均果实纵、横径分别为35.38 mm和29.21 mm。株高的变幅为4.4432.5 cm，变异系数和遗传多样性指数均较高，表明草莓资源的株高变异丰富。7肖桂林，等：73份草莓种质资源表型性状的遗传多样性分析及在湖北省的综合评价1551果树学报第40卷图 173 份栽培种草莓种质资源 34 个描述型性状变异类型频率分布及多样性指数Fig.1Frequency distribution and diversity index of 34 descriptive traits in 73 cultivated strawberry germplasms表 373 份栽培种草莓种质资源 17 个数量性状多样性分析Table 3Genetic diversity of 17 quantitative traits in 73 cultivated strawberry germplasms性状Trait株高Plant height,PH/cm冠径Canopy diameter,CD/cm叶片长Leaf length,LL/cm叶片宽Leaf width,LW/cm叶片长宽比Length-width ratio,LAR叶柄长度Petiole length,PL/cm叶柄粗度Petiole diameter,PD/mm复叶数量Number of compound leaves,CLN叶片厚Leaf thickness,Lthick/mm花梗长Pedicel length,PcelL/cm花梗粗Pedicel diameter,PcelD/mm花数Flower numbers,FN花冠径Flower diameter,FD/cm雄蕊数Stamen numbers,SN果质量Fruit weight,FW/g纵径Fruit longitudinal diameter,FLD/mm横径Fruit transverse diameter,FTD/mm均值标准差MeanSD17.076.3229.067.896.201.635.431.371.140.1213.695.592.780.6521.59.610.330.056.232.481.650.3828.4317.242.530.3526.463.4014.345.8935.386.1729.215.37变幅Range4.4432.5013.1655.503.0610.832.989.300.851.503.4829.961.524.954.8049.40.230.460.9013.2