A novel transcription factor FnMYB4 regulates pigments metabolism of yellow leaf mutants in Fragaria nilgerrensis

The strawberry species Fragaria nilgerrensis Schlechtendal ex J.Gay,renowned for its distinctive white,fragrant peach-like fruits and strong disease resistance,is an exceptional research material.In a previous study,an ethyl methane sulfonate(EMS)mutant library was established for this species,resulting in various yellow leaf mutants.Leaf yellowing materials are not only the ideal materials for basic studies on photosynthesis mechanism,chloroplast development,and molecular regulation of various pigments,but also have important utilization value in ornamental plants breeding.The present study focused on four distinct yellow leaf mutants:mottled yellow leaf(MO),yellow green leaf(YG),light green leaf(LG),and buddha light leaf(BU).The results revealed that the flavonoid content and carotenoid-to-chlorophyll ratio exhibited a significant increase among these mutants,while experiencing a significant decrease in chlorophyll and carotenoid contents compared to the wild type(WT).To clarify the regulatory mechanisms and network relationships underlying these mutants,the RNA-seq and weighted gene coexpression network(WGCNA)analyses were employed.The results showed flavonoid metabolism pathway was enriched both in MO and YG mutants,while the chlorophyll biosynthesis pathway and carotenoid degradation pathway were only enriched in MO and YG mutants,respectively.Subsequently,key structural genes and transcription factors were identified on metabolic pathways of three pigments through correlation analyses and quantitative experiments.Furthermore,a R2R3-MYB transcription factor,FnMYB4,was confirmed to be positively correlated with flavonoid synthesis through transient overexpression,virus-induced gene silencing(VIGS),and RNA interference(RNAi),accompanying by reoccurrence and attenuation of mutant phenotype.Finally,dual-luciferase(LUC)and yeast one-hybrid assays confirmed the binding of FnMYB4 to the FnFLS and FnF3H promoters,indicating that FnMYB4 positively regulates flavonoid synthesis.In addition,correlation analyses suggested that FnMYB4 also might be involved in chlorophyll and carotenoid metabolisms.These findings demonstrated the pivotal regulatory role of FnMYB4 in strawberry leaf coloration.

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。

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.

Strawberry (Fragaria spp.): Cultivation, Production, Consumption, and Marketing in Cameroon

Strawberry (Fragaria spp.) is one of the most important fruits classified as exotic fruits imported into Cameroon. To have an inventory of its cultivation in Cameroon, a survey study was carried out among eight farms of Fragaria spp. from January 2021 to February 2022. The plant was introduced in Cameroon in 2018. There are 13 varieties of Fragaria spp. currently cultivated. Among these 13 varieties, eleven are hybrids of Fragaria x ananassa (“Amiga”, “Amine”, “Camarosa”, “Chandler”, “Charlotte”, “Elsanta”, “Gariguette”, “Madame Moutot”, “Ostara”, “Ruby gem” and “San Andreas”), and two of the hybrids of Fragaria vesca (“Maestro” and “Mara des bois”). The cropping system, irrigation system, and type of fertilizers applied differ from one strawberry farm to another. Biofertilizers (such as mycorrhizal), inorganic and organic fertilizers are actually used to improve production. The potential annual production of strawberries from January 2021 to February 2022, estimated based on the survey data, was 21.216 tons for all growers. Among these eight production farms, the Lolodorf BIO Farm presents 6000 kg (six tons) of strawberries and 100,000 stolons (seedlings) produced, from seven varieties of Fragaria spp. cultivated, with 6 varieties which are hybrids variety Fragaria x ananassa (“Amiga”, “Amine”, “Chandler”, “Gariguette”, “Madame Moutot”, and “Ruby gem”), and one which is a hybrid of Fragaria vesca (“Mara des bois”). Certain diseases were also observed and recorded depending on the growing areas.

Antifungal Activity of Poncirus trifoliata Roots against Colletotrichum Species

Fruits and vegetables are an essential part of a healthy diet, providing humans with vitamins, phytonutrients, and minerals. They are significantly vulnerable, however, to post-harvest diseases caused by numerous fungal and bacterial pathogens. These pathogens can cause significant quantitative and qualitative losses from harvest to consumption during the handling and storage processes. Chemical fungicides are commonly used but are likely to leave residues on the produce, rendering short shelf-life produce, such as berries, unsuitable for human consumption. Identifying eco-friendly methods to control post-harvest disease is, therefore, of utmost importance. The presence of antifungal constituents in the roots of Poncirus trifoliata extracts was detected by thin layer chromatography-based bioautography. The active constituents were isolated and identified by bioautography assay-guided fractionation using flash chromatography followed by spectroscopic techniques. In this study, xanthoxyletin, demethylsuberosin, dentatin, nordentatin, ponfolin, and clausarin were isolated from the root extracts. The antifungal activity of these compounds was moderate to weak compared to the commercial fungicide captan. This study reports the isolation and identification of natural compounds from Poncirus trifoliata that exhibited antifungal activity against Colletotrichum fragariae and Botrytis cinerea, two major post-harvest pathogens.

基于改进Faster R-CNN模型的草莓果实识别算法

针对Faster R-CNN模型对自然状态下草莓(Fragaria ananassa Duch.)识别准确率不高的问题,以地垄种植草莓的实拍图片为数据源,采用改进RPN结构和更换主干特征提取网络的方法对Faster RCNN模型进行了改进。结果表明,改进Faster R-CNN模型识别成熟草莓平均精度(AP)为0.893 0,识别未成熟草莓平均精度(AP)为0.820 7,草莓识别准确率达到较高水平,解决了未成熟草莓识别困难的问题。同时,为了检验模型的自动计数性能,依据模型的识别结果建立了自动计数与人工计数的线性回归,成熟草莓、未成熟草莓的相关系数分别为0.973 7、0.944 7,自动计数与人工计数拥有较高的相关性,表明改进Faster R-CNN模型具有较高的识别性能与计数能力。

The telomere-to-telomere genome of Fragaria vesca reveals the genomic evolution of Fragaria and the origin of cultivated octoploid strawberry

Fragaria vesca,commonly known as wild or woodland strawberry,is the most widely distributed diploid Fragaria species and is native to Europe and Asia.Because of its small plant size,low heterozygosity,and relative ease of genetic transformation,F.vesca has been a model plant for fruit research since the publication of its Illumina-based genome in 2011.However,its genomic contribution to octoploid cultivated strawberry remains a long-standing question.Here,we de novo assembled and annotated a telomere-to-telomere,gap-free genome of F.vesca‘Hawaii 4’,with all seven chromosomes assembled into single contigs,providing the highest completeness and assembly quality to date.The gap-free genome is 220785082 bp in length and encodes 36173 protein-coding gene models,including 1153 newly annotated genes.All 14 telomeres and seven centromeres were annotated within the seven chromosomes.Among the three previously recognized wild diploid strawberry ancestors,F.vesca,F.iinumae,and F.viridis,phylogenomic analysis showed that F.vesca and F.viridis are the ancestors of the cultivated octoploid strawberry F.×ananassa,and F.vesca is its closest relative.Three subgenomes of F.×ananassa belong to the F.vesca group,and one is sister to F.viridis.We anticipate that this high-quality,telomere-to-telomere,gap-free F.vesca genome,combined with our phylogenomic inference of the origin of cultivated strawberry,will provide insight into the genomic evolution of Fragaria and facilitate strawberry genetics and molecular breeding.

Rapid evolution of T2/S-RNase genes in Fragaria linked to multiple transitions from self-incompatibility to self-compatibility

The T2/RNase gene family is widespread in eukaryotes,and particular members of this family play critical roles in the gametophytic self-incompatibility(GSI) system in plants.Wild diploid strawberry(Fragaria)species have diversified their sexual systems via self-incompatible and self-compatible traits,yet how these traits evolved in Fragaria remains elusive.By integrating the published and de novo assembled genomes and the newly generated RNA-seq data,members of the RNase T2 gene family were systematically identified in six Fragaria species,including three self-incompatible species(Fragaria nipponica,Fragaria nubicola,and Fragaria viridis) and three self-compatible species(Fragaria nilgerrensis,Fragaria vesca,and Fragaria iinumae).In total,115 RNase T2 genes were identified in the six Fragaria genomes and can be classified into three classes(Ⅰ-Ⅲ) according to phylogenetic analysis.The identified RNase T2 genes could be divided into 22 homologous gene sets according to amino acid sequence similarity and phylogenetic and syntenic relationships.We found that extensive gene loss and pseudogenization coupled with small-scale duplications mainly accounted for variations in the RNase T2 gene numbers in Fragaria.Multiple copies of homologous genes were mainly generated from tandem and segmental duplication events.Furthermore,we newly identified five S-RNase genes in three self-incompatible Fragaria genomes,including two in F.nipponica,two in F.viridis,and one in F.nubicola,which fit for typical features of a pistil determinant,including highly pistil-specific expression,highly polymorphic proteins and alkaline isoelectric point(pI),while no S-RNase genes were found in all three selfcompatible Fragaria species.Surprisingly,these T2/S-RNase genes contain at least one large intron(>10 kb).This study revealed that the rapid evolution of T2/S-RNase genes within the Fragaria genus could be associated with its sexual mode,and repeated evolution of the self-compatible traits in Fragaria was convergent via losses of S-RNase.

氟吡呋喃酮等六种杀虫剂对草莓蚜虫的田间防效

通过3年田间试验,比较了200 g/L氟吡呋喃酮SL、240 g/L螺虫乙酯SC、10%氟啶虫酰胺WG、50 g/L双丙环虫酯DC、70%吡虫啉WG和22%氟啶虫胺腈SC对草莓(Fragaria ananassa Duch.)蚜虫的防治效果,以及200 g/L氟吡呋喃酮SL不同施药量对草莓蚜虫的田间防效。结果表明,供试6种杀虫剂对草莓蚜虫均有较好的防治效果,其中200 g/L氟吡呋喃酮SL、10%氟啶虫酰胺WG和50 g/L双丙环虫酯DC防治效果更突出,速效性好、持效期长;200 g/L氟吡呋喃酮SL对草莓蚜虫的防治效果较好,建议施药量在90.0~120.0 g a.i./hm^(2)为宜。

草莓褐色轮斑病病原鉴定及室内药效研究

草莓褐色轮斑病近年越来越严重,尤其在草莓育苗阶段。从草莓发病叶片、匍匐茎上分离得到病原菌,对病原菌进行形态特征观察、生物学特性研究、ITS序列分析以及室内药效试验。结果表明:该病原菌为Sphae-ronaemella fragariae。该菌菌丝生长最适温度范围是25～28℃;适宜pH为6;在供试的几种碳、氮源中,最适的碳源是蔗糖,最适的氮源是酵母浸出液。在供试的9种药剂中,以咪鲜胺1 000倍液对病菌的抑制作用最好。

草莓根部侵染草莓炭疽菌的致病动态变化过程

为了从组织学和细胞学水平上了解草莓炭疽菌(Colletotrichum fragariae)病原菌致病机理及其与寄主的互作机制,通过草莓炭疽菌分生孢子液接种侵染健康草莓幼苗根,在荧光显微镜下观察病原菌的侵染过程,研究草莓根部浸染草莓炭疽菌的致病性。结果表明,接种12 h,95%的分生孢子萌发;接种24 h,69%芽管顶端产生附着胞或附着枝并形成侵入钉开始侵染根表皮细胞;接种48 h,产生大量菌丝并纠结成网状;接种4 d,少量的根尖开始变褐;接种9 d,整个根系几乎变为黑褐色,产生少量分生孢子盘;接种13 d,侵染菌丝扩展到主轴根微管柱,新的分生孢子产生,完成一个侵染循环;接种20 d,侵染菌丝入侵到达叶柄和花茎;接种41 d,侵染菌丝扩散到叶柄表面并侵染叶柄腺毛;接种55 d,侵染菌丝入侵到达叶片并侵染叶片腺毛,组织细胞开始崩解死亡,叶片上形成典型的炭疽病病斑。[结论]:C.fragariae能通过根侵染到整株植株,属于系统性侵染。

草莓枯萎病菌的分离与鉴定

从湖北省武汉市郊区草莓种植户的大棚内采集萎蔫的植株,分离致病菌,采用PCR扩增、病原菌形态鉴定、致病性、分子检测等方法检测草莓枯萎病菌。结果表明,形态观察为尖孢镰刀菌(Fusarium oxysporum),接种草莓后出现典型的草莓枯萎病菌症状;采用真菌的通用引物对ITS1和ITS4进行PCR检测,分离得到的致病菌株均能扩增出分子量为567 bp的特异性条带。结合形态、致病性测定和分子检测,确定分离和鉴定的病原菌为草莓枯萎病菌(Fusarium oxysporum Schlecht.f.sp.fragariae Winks et Williams)。

湖北省草莓育苗期炭疽病病害调查

2012年夏季,对湖北省草莓(Fragaria ananassa)育苗期炭疽病发病情况进行了调查,主要调查对象为湖北省草莓主栽品种晶瑶、丰香、法兰地等。结果表明,晶瑶育苗期炭疽病发病情况最严重,平均发病率为41.7%,平均病情指数为21.5;丰香次之;法兰地发病最轻,平均发病率为16.2%,平均病情指数为7.5。并对炭疽病病样进行了分离和鉴定。

溶菌酶和乳酸链球菌素在草莓保鲜中的应用研究

为了研究溶菌酶和乳酸链球菌素(Nisin)对草莓(Fragaria ananassa Duch.)的低温保鲜效果,使用不同浓度的溶菌酶与Nisin复配液对草莓进行保鲜处理,贮藏于4℃,对贮藏期间草莓的烂果率、失重率、维生素C含量、还原糖含量、总酸含量进行测定。结果表明,0.01%溶菌酶与0.10%Nisin复配保鲜液对草莓保鲜效果最好,贮藏前5 d无烂果。贮藏第五天草莓失重率为8.24%;维生素C含量为109.0 mg/100 g,下降13.6%;还原糖含量为7.21 g/100 g,上升14.3%;总酸含量为6.21 g/kg,下降43.1%。

钙对干旱胁迫下草莓SOD、CAT、PPO活性的影响

采用自然干旱胁迫的方法,以‘章姬’、‘丰香’为材料,研究了干旱胁迫下不同Ca2+浓度处理对草莓(Fragaria Ananassa Duchesne)叶片超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、多酚氧化酶(PPO)的影响。结果表明,在干旱胁迫下,外源钙能在一定程度上维持酶的活性,以减轻活性氧对草莓的伤害,从而在一定程度上提高草莓抗旱性,且不同处理之间存在差异,以10mmol/LCaCl2处理效果最好。

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。

不同复合基质对‘红颊’草莓产量和品质的影响

[目的]研究6种不同复合基质对‘红颊’草莓营养生长、物质分配、产量和品质的影响。[方法]测定不同基质栽培下‘红颊’草莓植株的形态指标、植株鲜(干)重、产量以及果实可溶性固形物、总酸、维生素C、总糖、还原糖和蔗糖含量。[结果]不同基质栽培对‘红颊’草莓植株的形态指标影响较小,对植株鲜(干)重的影响较大,且各处理间产量和品质指标差异明显。综合评价,该试验中以泥炭∶砻糠灰∶珍珠岩=3∶1∶1(V/V/V)的复合基质栽培效果最好。[结论]复合基质泥炭∶砻糠灰∶珍珠岩=3∶1∶1适合‘红颊’草莓生长,值得在草莓基质栽培中应用。

草莓新品种晶玉对炭疽病和灰霉病的抗性鉴定

采用田间试验和人工接种的方法对‘晶玉’、‘晶瑶’、‘丰香’和‘甜查理’的炭疽病以及灰霉病抗性进行了鉴定和比较。结果表明,草莓(Fragaria ananassa Duch.)新品种‘晶玉’对炭疽病具有中等抗性水平,其抗性水平低于‘甜查理’,但高于‘晶瑶’和‘丰香’;‘晶玉’对灰霉病具有一定的耐病性,具有花不易感灰霉病和果实发病率显著低于‘甜查理’的优点。

草莓花枯病原真菌的PCR检测

为了建立简易、快速的草莓花枯病原真菌(Rhizoctonia fragariae)的PCR检测技术,以从美国ATCC购买的标准菌株14691TM为试验材料,通过真菌的通用引物对ITS4和ITS5的PCR扩增,从受感染草莓的根、叶、花、果实4个部位分离的菌株扩增到长度为670 bp的特异性条带,DNA测序证实了这一扩增产物与NCBI的GenBank内草莓花枯病原真菌的相关序列有高度序列同源性。结合草莓根、叶、花、果实的接种及分离、病原菌形态观察及致病性鉴定等结果,确定通用引物PCR扩增技术可以作为草莓花枯病菌检测的基本方法。

武汉市城郊草莓产业发展情况调研报告

对武汉市城郊草莓(Fragaria ananassa Duch.)主要产区的2014年种植情况进行了调查研究。结果表明,2014年度武汉市黄陂区草莓种植面积为780 hm^2,东西湖区草莓种植面积为487 hm^2;栽培模式为促成栽培,经济效益显著。存在的问题主要是高温病害多,农药使用量较大,食品安全问题突出;抗病优质品种缺乏仍是技术瓶颈,新技术普及率低。建议找准市场定位,增加科技投入,拓展草莓产业空间,提高组织化程度,增强市场竞争力。