Genome-wide characterization and expression analysis of WRKY family genes during development and resistance to Colletotrichum fructicola in cultivated strawberry(Fragaria×ananassa Duch.)

Based on the recently published whole-genome sequence of cultivated strawberry ’Camarosa’, in this study, 222FaWRKY genes were identified in the ’Camarosa’ genome. Phylogenetic analysis showed that the 222 FaWRKY candidate genes were classified into three groups, of which 41 were in group Ⅰ, 142 were in group Ⅱ, and 39 were in group Ⅲ. The 222 FaWRKY genes were evenly distributed among the seven chromosomes. The exon–intron structures and motifs of the WRKY genes had evolutionary diversity in different cultivated strawberry genomes. Regarding differential expression, the expression of FaWRKY133 was relatively high in leaves, while FaWRKY63 was specifically expressed in roots. FaWRKY207, 59, 46, 182, 156, 58, 39, 62 and 115 were up-regulated during achene development from the green to red fruit transition. FaWRK181, 166 and 211 were highly expressed in receptacles at the ripe fruit stage. One interesting finding was that Fa WRKY179 and 205 were significantly repressed after Colletotrichum fructicola inoculation in both ’Benihoppe’ and ’Sweet Charlie’ compared with Mock. The data reported here provide a foundation for further comparative genomics and analyses of the distinct expression patterns of FaWRKY genes in various tissues and in response to C. fructicola inoculation.

The transcriptional landscape of cultivated strawberry(Fragaria×ananassa)and its diploid ancestor(Fragaria vesca)during fruit development

Cultivated strawberry(Fragaria×ananassa)originated from four diploid ancestors:F.vesca,F.viridis,F.iinumae and F.nipponica.Among them,F.vesca is the dominant subgenome for cultivated strawberry.It is not well understood how differences in gene expression between diploid and octoploid strawberry contribute to differences during fruit development.In this study,we used comprehensive transcriptomic analyses of F.vesca and F.×ananassa to investigate gene expression at the different stages of fruit development.In total,we obtained 3508(turning stage)and 3958(red stage)differentially expressed genes with pairwise comparisons between diploid and octoploid.The genes involved in flavonoid biosynthesis were almost upregulated in the turning stages of octoploid,and we also discovered a ripe fruit-specific module associated with several flavonoid biosynthetic genes,including FveMYB10,FveMYB9/11,and Fve RAP,using weighted gene coexpression network analysis(WGCNA).Furthermore,we identified the species-specific regulated networks in the octoploid and diploid fruit.Notably,we found that the WAK and F-box genes were enriched in the octoploid and diploid fruits,respectively.This study elucidates new findings on flavonoid biosynthesis and fruit size of strawberry with important implications for future molecular breeding in cultivated strawberry.

Study on Ornamental Cultivation Techniques of Strawberry(Fragaria×ananassa Duch)

Strawberry(Fragaria×ananassa Duch)is a very popular fruit,which is very common for productive cultivation,but not for ornamental cultivation.In order to improve the ornamental quality of strawberry and study its ornamental cultivation techniques,the variety selection,cultivation methods,flower pot selection,soil,fertilizer and water management,temperature,humidity and light management,and flower,fruit and pollination management of strawberry were investigated,especially its precision fertilization.The optimum combination fertilization scheme was A_(3)B_(2)C_(1),namely‘Lvbao’6 g,‘Weibao’6 g,‘Double carbon’2 g,which was obtained by an orthogonal test with three factors and three levels.This combined fertilization method overcomes the deficiency of single fertilization and improves the fertilization effect,and provides a reference for ornamental cultivation of strawberry.

草莓(Fragaria ananassa Duch)茎段组织培养的研究

实验在草莓茎段的组织培养过程中,对外植体不同消毒时间,最佳诱导培养基、分化培养基与生根培养基进行了筛选。结果表明,0.1%升汞对茎段消毒2min时效果最好;诱导愈伤的最佳培养基为:MS+1.5mg·L-16-BA+0.1mg·L-1NAA;最佳分化的培养基为:MS+2.0mg·L-16-BA+0.2mg·L-1NAA;最佳生根培养基为:MS+0.25mg·L-16-BA。

5-Aminolevulinic acid against strawberry Fusarium wilt:Bidirectional regulation of biocontrol agents and pathogens

Strawberry Fusarium wilt (SFW) is a systematic soil-borne disease caused by Fusarium oxysporum f.sp.fragaria (Fof),which infects the vascular bundles,blocking water and nutrient transport from roots to the aboveground.It is a severe pathogen which spreads rapidly and destroys strawberry production.Finding a way to control this disease is of great scientific value and practical importance.In this study,three fungi were isolated from the vascular tissues of sick strawberries in the field.After DNA sequencing,they were identified as Fof,Aspergillus fumigatus and Trichoderma harzianum,respectively,among which the first two are pathogens and the third is a probiotic.All fungi were controlled by thiophanate-methyl (TM),a commercial fungicide.On PDA medium,20 mg·L^(-1)5-aminolevulinic acid (ALA),a natural non-protein amino acid,promoted T.harzianum proliferation,but inhibited Fof and A.fumigatus.In confrontation test,the growth of Fof or A.fumigatus was inhibited by T.harzianum and exogenous ALA promoted T.harzianum growth but significantly inhibited the pathogen growth.When three species of fungi were separately or combinedly inoculated on healthy strawberry plants,T.harzianum promoted plant growth and development while Fof or A.fumigatus caused growth retardation,where Fof directly caused leaf yellowing and plant wilting.When the plants inoculated with different fungus were treated with ALA,the results turned out that ALA alleviated SFW symptoms by bidirectionally promoting T.harzianum proliferation and inhibiting Fof and A.fumigatus.Thus,ALA might be used in comprehensively controlling SFW in strawberry industry.

中国草莓属(Fragaria)植物的分类研究

中国是世界上野生草莓种类最丰富的国家。近20年来,从国内外收集保存了103份野生草莓资源,鉴定出我国自然分布有11个种,约占世界草莓属植物20个种的一半。这11个种包括8个二倍体种:森林草莓(Fragaria vescaL.)、黄毛草莓(F.nilgrrensisSchlecht.)、五叶草莓(F.pentaphyllaLozin-sk.)、纤细草莓(F.gracilisLozinsk.)、西藏草莓(F.nubicolaL ind l.)、绿色草莓(F.viridisDuch.)、裂萼草莓(F.daltonianaGay)、东北草莓(F.mandschuricaStaudt)和3个四倍体种:东方草莓(F.oriental-isLozinsk.)、西南草莓〔F.m oupinensis(Franch.)Card.〕、伞房草莓(F.corym bosaLozinsk.)。对我国原产野生草莓种类进行了较系统的性状描述和分类研究,并列出了我国野生草莓分种检索表。提出我国分布有11个种的结论比以前记载的8个种更全面和完整。

Cloning and Sequencing the Full-length cDNA of Annexin from Strawberry Fruit

采用RACE技术从草莓 (FragariaananassaDuch .)成熟果实中分离克隆了膜联蛋白 (annexin)基因的cDNA 5′_端未知序列 ,并通过测序确定了翻译的起始密码位点、终止密码位点及完整的读码框 ,从而首次获得了草莓果实膜联蛋白基因的cDNA全序列 ,命名为annfaf(annexinofFragariaananassafruit)基因。

新疆天山野生草莓与绿色草莓(Fragariaviridis Duch.)同一性的鉴定

通过植物学性状观察和RAPD技术鉴定证明新疆天山野生草莓为绿色草莓 (Fra gariaviridisDuch .) ,表明中国自然分布的草莓野生种达到 9个。绿色草莓在分类学上的典型特征为匍匐茎上从第二节开始每节均形成幼苗 ;花序直立、高于叶面 ,花瓣显露时呈黄绿色 ,雄蕊花丝细长 ,高于雌蕊 ;果实圆形或扁圆形 ,淡绿色 ,阳面略红 ,果硬 ,成熟时萼片紧贴 ,脱萼难。

草莓属二倍体种东北草莓(Fragariamandschurica Staudt)研究

从中国吉林、黑龙江、内蒙古收集到5份形态相近的二倍体野生草莓,经多年植物分类学性状观察鉴定为东北草莓(FragariamandschuricaStaudt),该中文名为首次赋予,此前国内尚无该种的记载。“Manchuria”意为满州,是对中国东北的旧称。该种植株绒毛多少及其着生方式、花器官、种子等诸多性状明显不同于森林草莓(F.vescaL.)。RAPD(RandomAmplifiedPolymorphicDNA)鉴定技术也对这一结果给予有力支持,而且表明二倍体东北草莓与四倍体东方草莓(F.orientalisLozinsk)、六倍体麝香草莓(F.moschataDuch.)的亲缘关系最近,东北草莓可能是它们的原始种。

Exogenous Melatonin Improves Physiological Characteristics and Promotes Growth of Strawberry Seedlings Under Cadmium Stress

Melatonin,as a plant growth regulator,is involved in plant stress resistance.We studied the effects of different concentrations(0,10,50,100,150,and 200μmol·L−1)of melatonin on the growth and physiological characteristics of strawberry under cadmium(Cd)stress.The results represented that the growth of strawberry seedlings was inhibited under Cd stress,and the seedling biomass,chlorophyll content and the activities of antioxidant enzymes such as superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),and ascorbate peroxidase(APX)decreased.These toxic effects were,however,effectively remediated by exogenous melatonin pretreatment.Melatonin treatment slowed the inhibitory effect of seedling growth and significantly increased seedling biomass,chlorophyll content,antioxidant enzyme activities,and soluble protein levels in leaves and roots compared with the control.Melatonin treatment also reduced the malondialdehyde(MDA)content and the oxidative stress reactions,increased the anthocyanin content,and slowed the senescence rate,thereby reducing the toxic effects of Cd on strawberries.This indicates that foliar application of melatonin can effectively remediate the adverse effects of Cd on strawberry seedlings;the greatest effect was obtained with melatonin concentration of 100μmol·L−1.

Analysis of Volatile Components in Strawberry Cultivars Xingdu 1 and Xingdu 2 and Their Parents

To study differences and genetic trends of volatiles in Xingdu 1 and Xingdu 2, the strawberry cultivars, with their parents, the volatile components were analyzed by head space-solid phase microextraction and gas chromatography-mass spectrometry. The results indicated that a total of 50 compounds were identified in Xingdu 1 cultivar, including hexanoic acid methyl ester and hexanoic acid ethyl ester, butanoic acid methyl ester, and butanoic acid ethyl ester, octanoic acid methyl ester and octanoic acid ethyl ester, isomyl hexanoate. And there were 71 compounds in Xingdu 2, which were similar to that of Xingdu 1. About 55 volatiles were identified in Allstar （female parent）, including acetone, acetic acid 1- methylethyl ester, 1-octanol, hexanoic acid ethyl ester and butanoic acid methyl ester. And 43 volatiles were identified in Toyonoka （male parent）, including hexanoic acid ethyl ester, nerolidol, 1-octanol, hexanoic acid methyl ester, acetic acid octyl ester, isomyl hexanoate and octanoic acid methyl ester. Consequently, hexanoic acid methyl ester and hexanoic acid ethyl ester, butanoic acid methyl ester, and butanoic acid ethyl ester and isomyl hexanoate in twins might be affected both sides of their parents. In the family, male parent （Toyonoka） had the highest concentrations of fragrance alcohols, such as nerolidol, linalool and 1-octanol, but which couldn＇t be transmitted to the descendants. Acetone, acetic acid 1- methylethyl ester of the twins originally came from matrilinear inheritance, as the contents of the two compounds were very high in Allstar but little in Toyonoka. Genetic trends of butanoic acid methyl ester, butanoic acid ethyl ester, hexanoic acid ethyl ester, hexanoic acid methyl ester and isomyl hexanoate were affected by both sides of parents.

Root Growth, Mycorrhizal Frequency and Soil Microorganisms in Strawberry as Affected by Biopreparations

The aim of the study was to assess the effects of various biopreparations on the growth of the strawberry root system, the number of spores of mycorrhizal fungi, the total number of bacteria in the rhizosphere soil, and the degree of mycorrhizal association in the roots of two strawberry cultivars. The experiment with strawberry plants was established in the spring of 2010 in the Experimental Orchard of the Institute of Horticulture in Dabrowice. The objects of research were “frigo” strawberry plants of the cultivars Elsanta and Elkat. The following experimental combinations were used: control, control NPK (standard NPK fertilization), manure, mycorrhizal preparation Micosat F, Humus UP, Humus Active + Aktywit PM, BioFeed Amin, BioFeed Quality, Tytanit, Vinassa, Florovit Eko, and Florovit Pro Natura. The use of the biopreparation BioFeed Quality resulted in a six-fold increase in root length and a seven-fold increase in root surface area. Compared with NPK fertilization, application of the preparation BioFeed Amin contributed to an eight-fold increase in root volume, and the use of Vinassa increased 24-fold the number of root tips of Elkat strawberry plants. Micosat F and Humus UP caused a five-fold increase in mycorrhizal frequency in the roots of strawberry plants. Micosat F and manure contributed to a two- and four-fold increase, respectively, in the number of spores in the rhizosphere soil. Application of the preparations Humus UP, BioFeed Amin and Florovit Eko doubled the total number of bacteria and filamentous fungi in the rhizosphere soil of strawberry plants of the cultivars Elsanta and Elkat in comparison with NPK fertilization. Fertilization with the biopreparations intensified the growth of the root system and increased the number of spores of AM fungi, mycorrhizal frequency, and the total number of bacteria and filamentous fungi in the soil.

Differential Pathogenic Response in Strawberry Tissues and Organs by Colletotrichum acutatum

The susceptibility of different tissues and organs from strawberry plants, cv ＂Camarosa＂, to Colletotrichum acutatum was tested using a severity index based on infection response. Symptoms developed on inoculated tissues were characterized along 30 days. Flowers, except sepals, petioles and fruits were the most susceptible organs to the pathogen and they became necrotic tissues at 30 days post inoculation （dpi）. Also, well-developed acervuli, which produced masses of orange-pink spores, were observed on these infected organs. An asymptomatic stage or latency phase was observed in green and white strawberry fruits. In spite of they were inoculated anthracnose symptoms were observed only when they became red fruits. On the other hand, strawberry leaves and sepals were resistant to infection by C. acutatum and only small flecks or light brown spots were observed reaching a size of 1 to 5 mm at 30 dpi. Likewise, the susceptibility of stolons and crowns to C. acutatum was evaluated as intermediate at 30 dpi. Finally, the infection process of the fungus on strawberry leaves and petioles was studied using light and electron microscopy. Pre-penetration events were similar on both, leaves and petioles： However, differences between colonization of strawberry leaves and petioles by C. acutatum were observed.

DNA电击对草莓花粉活力的影响

以‘申琪’草莓花粉为材料,结合不同电脉冲形态、电场强度和电击次数进行处理,然后在花粉萌发培养基上28℃黑暗孵育,于3、6、18、24h时用显微镜观察花粉萌发情况,探索草莓花粉电击适宜参数。结果表明:80μs方波脉冲在4.5—6.0kV/cm强度电击1次,可逆抑制花粉萌发,抑制程度与电场强度呈正相关;电击后3h花粉萌发率为CK1的36%—70%,24h时为CK1的60%—85%。方波4.5—5.0kV/cm强度电击2次产生部分可逆抑制,电击24h后花粉萌发率为CK1的20%—50%;5.5—6.0kV/cm方波脉冲电击2次和4.5—6.0kV/cm电击3次产生不可逆抑制,电击24h后花粉萌发率低于CK1的10%。用2.75—6.0kV/cm指数衰减波3.5ms脉冲电击后,花粉活力快速下降,电击18h后花粉萌发率在1%以下。综上,强度4.5—5.0kV/cm方波脉冲80μs电击2次为草莓花粉适宜电击参数。

柠檬醛对草莓贮藏期间品质和抗氧化能力的影响

目的探讨不同植物精油对草莓贮藏期间品质和抗氧化能力的影响。方法以“红颜”草莓为试材,分别用不同质量浓度(40、80、160和320μg/mL)的百里香精油、丁香花精油、肉桂精油、枯茗醛和柠檬醛进行处理,测定其在20℃贮藏期间的失重率、腐烂指数、理化性质、抗氧化物质含量和抗氧化酶活性等指标。结果柠檬醛在40μg/mL质量浓度时具有最佳保鲜效果,能够有效地降低草莓果实的失重率和腐烂指数,延缓可溶性固形物、可滴定酸和抗坏血酸含量的下降,维持较高的总酚和总黄酮含量,并提高果实对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基和羟基自由基的清除能力。此外,该处理还能显著提高过氧化氢酶、超氧化物歧化酶、抗坏血酸过氧化物酶等抗氧化酶的活性,且有效抑制丙二醛含量的升高。结论柠檬醛在草莓贮藏中具有良好的保鲜效果,能够保持其品质和营养价值,这为柠檬醛在水果贮藏保鲜中的应用提供了依据。

‘红颜’草莓组培快繁技术试验

以‘红颜’草莓匍匐茎的茎尖为试材,经初代培养和继代培养获得脱毒苗。结果表明:初代愈伤诱导培养最适宜的培养基配方为MS+NAA 1.0 mg·L^(-1)+6-BA 5 mg·L^(-1);最适宜愈伤增殖的培养基配方为MS+NAA 1 mg·L^(-1)+6-BA 10 mg·L^(-1)+TDZ 1 mg·L^(-1);最适宜诱导萌芽的培养基配方为MS+NAA 0.40 mg·L^(-1)+6-BA 10 mg·L^(-1)+活性炭1g·L^(-1)。在继代培养中最适宜诱导愈伤的部位为叶片,最适宜诱导萌芽的部位为茎尖处;最适宜叶片诱导出愈伤组织的培养基配方为MS+NAA 0.50 mg·L^(-1)+TDZ 20 mg·L^(-1)+活性炭1g·L^(-1);最适宜茎尖诱导萌芽的培养基配方为MS+IBA 5 mg·L^(-1)+活性炭3 g·L^(-1);最佳生根培养基配方为1/2MS+IBA 5 mg·L^(-1)+活性炭3 g·L^(-1)。

设施草莓病虫害发生特点及绿色防控技术

由于设施环境的封闭性和湿度较高,病虫害呈多样化且易发生,严重影响设施草莓(Fragaria×ananassa Duch.)栽培的产量和品质。为便于设施草莓种植者及时发现和诊断病虫害发生情况,对设施草莓病虫害发生特点、影响因素、为害部位等进行梳理分析,同时提出通过优化栽培措施,结合物理和生物防治手段,辅以高效、低毒、低残留的化学农药等构建设施草莓病虫害绿色防控体系,以期为设施草莓栽培产业的绿色高效发展提供参考。

合肥草莓产业发展现状及对策建议

在介绍合肥草莓产业发展现状、主要做法的基础上,分析了草莓产业发展中存在的问题,提出了相应的解决对策。

基于果形对草莓品种遗传多样性SSR分析

本研究利用SSR分子标记技术对国家果树种质北京草莓圃的173份草莓品种进行区分,根据果形利用PopGene软件对其做了聚类分析。结果显示,长圆锥形、短圆锥形、阔圆锥形、圆锥形/长楔形、圆锥形/平楔形、长楔形、近圆形、圆球形/卵圆形的观测纯合度都为0.846 2,扁球形、圆球形/半圆球形、短圆锥形/短楔形的观测纯合度都在0.923 1~0.948 7之间,但是短圆形纯合度最低,为0.692 3,由此发现与其他形状的亲缘关系较远,其余形状相互之间的亲缘关系较近。根据果形分析SSR扩增等位基因的多态性发现,173份草莓品种整体纯合体偏多,其中楔形/长圆锥形的无杂合体,而短圆形的纯合体最少,草莓品种整体亲缘关系较近。

富氢水对提高温室高架基质栽培草莓品质及抗逆性的效果

为了研究富氢水(Hydrogen-rich water,HRW)对草莓生长的影响,以“法兰地”草莓为材料,采用HRW和自来水分别浇灌草莓根部,通过对果实品质指标可溶性固形物、可溶性糖含量、可滴定酸含量、总黄酮含量、总酚含量、可溶性蛋白含量、硝酸盐含量和花青素含量进行测定,并分别对根系、叶片的抗逆指标丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性、过氧化物酶(POD)活性、抗坏血酸过氧化物酶(APX)活性、脯氨酸(Pro)含量进行测定,分析灌溉HRW对温室高架基质栽培草莓的果实品质以及对植株抗逆能力的影响。结果表明:HRW灌溉的草莓果实可溶性糖含量、可滴定酸含量、总黄酮含量、总酚含量及根系CAT活性、APX活性分别比自来水对照显著提升6.5%、8.4%、7.9%、4.0%及7.9%、7.4%,叶片SOD活性极显著提升23.4%,叶片Pro含量和根系MDA含量、SOD活性、POD活性分别极显著降低8.3%、8.4%、11.9%、18.3%。说明HRW灌溉处理可提升草莓抗氧化能力,并通过调节抗氧化酶活性的高低参与草莓根系抗逆性调控,从而提高草莓的口感和品质,达到提质增效的目的。