Advances in Cotton Breeding for Resistance to Fusarium and Verticillium Wilt in the Last Fifty Years in China

This review summarizes the main advances in cotton breeding for resistance to fusarium and verticillium wilt in the last fifty years in China. The topics discussed include main achievements, experiences, problems and countermeasures for solving.

Test on Wilt-Resistance of Simple Hybrides F7 and Beckross Hybrid Lines F6 B1 of <i>G. hirsutum</i>L. of Cotton to New Virulent Isolates of Fungus <i>Fusarium verticillioides</i>and <i>Verticillium dahliae</i>

The article presents the results of studies on the resistance of hybrid cotton lines to a new virulent isolate (strain) of the fungus Fusarium verticillioides upon inoculation of the host plant. Based on the studies, it was found that the complex genotypic resistance of the studied lines, when the host plants are inoculated with isolates of -100 V. dahliae Kleb fungus and 103 Fusarium verticillioides fungi, depends on the degree of resistance of the parental forms and their combination ability.

Characterization of salt tolerance and Fusarium wilt resistance of a sweetpotato mutant

The variant LM1 was previously obtained using embryogenic cell suspension cultures of sweetpotato variety Lizixiang by gamma-ray induced mutation, and then its characteristics were stably inherited through six clonal generations, thus this mutant was named LM1. In this study, systematic characterization of salt tolerance and Fusarium wilt resistance were performed between Lizixiang and mutant LM1. LM1 exhibited significantly higher salt tolerance compared to Lizixiang. The content of proline and activities of superoxide dismutase（SOD） and photosynthesis were significantly increased, while malonaldehyde（MDA） and H_2O_2 contents were significantly decreased compared to that of Lizixiang under salt stress. The inoculation test with Fusarium wilt showed that its Fusarium wilt resistance was also improved. The lignin, total phenolic, jasmonic acid（JA） contents and SOD activity were significantly higher, while H_2O_2 content was significantly lower in LM1 than that in Lizixiang. The expression level of salt stress-responsive and disease resistance-related genes was significantly higher in LM1 than that in Lizixiang under salt and Fusarium wilt stresses, respectively. This result provides a novel and valuable material for improving the salt tolerance and Fusarium wilt resistance of sweetpotato.

Development and identification of Verticillium wilt-resistant upland cotton accessions by pyramiding QTL related to resistance

Cotton Verticillium wilt is a serious soil-borne disease that leads to significant losses in fiber yield and quality worldwide. Currently, the most effective way to increase Verticillium wilt resistance is to develop new resistant cotton varieties. Lines 5026 and 60182 are two Verticillium wilt-resistant upland cotton accessions. We previously identified a total of 25 quantitative trait loci（QTLs） related to Verticillium wilt resistance from 5026 and 60182 by assembling segregating populations from hybridization with susceptible parents. In the current study, using 13 microsatellite markers flanking QTLs related to Verticillium wilt resistance, we developed 155 cotton inbred lines by pyramiding different QTLs related to Verticillium wilt resistance from a filial generation produced by crossing 5026 and 60182. By examining each allele＇s effect and performing multiple comparison analysis, we detected four elite QTLs/alleles（q-5/NAU905-2, q-6/NAU2754-2, q-8/NAU3053-1 and q-13/NAU6598-1） significant for Verticillium wilt resistance, pyramiding these elite alleles increased the disease resistance of inbred lines. Furthermore, we selected 34 elite inbred lines, including five lines simultaneously performing elite fiber quality, high yield and resistance to V. dahliae, 14 lines with elite fiber quality and disease resistance, three lines with high yield and disease resistance, and 12 lines with resistance to V. dahliae. No correlation between Verticillium wilt resistance and fiber quality traits/yield and its components was detected in the 155 developed inbred lines. Our results provide candidate markers for disease resistance for use in marker-assisted breeding（MAS）, as well as elite germplasms for improving important agronomic traits via modern cotton breeding.

QTL Analysis of Fiber Yield and Quality and Resistance to Verticillium Wilt Using Gossypium hirsutum and G.barbadense Advanced Backcross Populations

To introgress elite QTL alleles of Gossypium barbadense L.for fiber yield and quality and resistance to Verticillium wilt into G.hirsutum L.,enlarge the genetic base of G.hirsutum,and

Studies on Cotton Breeding Resistant to Fusarium and Verticillium wilt Diseases

Both Fusarium and Verticillium wilts are important soil-borne diseases,which can not be effectively controlled by chemical fungicides.The two diseases,especially Verticillium wilt,have

Induction of Tolerance to Fusarium Wilt and Defense-Related Mechanisms in the Plantlets of Susceptible Berangan Banana Pre-Inoculated with Pseudomonas sp.(UPMP3) and Burkholderia sp.(UPMB3)

This study is aimed at assessing the ability of two endophytic bacteria originally isolated from healthy oil palm roots, Pseudomonas sp. （UPMP3） and Burkholderia sp. （UPMB3） to induce resistance in susceptible Berangan banana against Fusarium oxysporum race 4 （FocR4）. Increased accumulation of resistance-related enzymes such as peroxidase （PO）, phenylalanine ammonia lyase （PAL）, lignithioglycolic acid （LTGA）, and pathogenesis-related （PR） proteins （ehitinase and β-1,3-glucanase） has been observed in plantlets treated with endophytic bacteria UPMP3 and UPMB3 singly or as mixture under glasshouse conditions. Pre-inoculation of banana plantlets with UPMP3 showed a significant reduction in Fusarium wilt incidence 72 d after challenged inoculation with FocR4. UPMB3 was less effective in suppressing Fusarium wilt compared to UPMP3, whereas, the mixture of both endophytes showed an intermediate effect. Based on these results, it is concluded that UPMP3 could be a promising biological control agent that can trigger resistance against Fusarium wilt in susceptible Berangan banana.

Quantitative Trait Loci for Resistance Against Fusarium Wilt Based on Three Cotton F_2 Populations

Fusarium wilt （FW） is one of the most common cotton diseases in the world. Identification of QTLs conferring resistance to FW is key for the incorporation of resistance genes into elite cultivars. Two intraspecific （cross between Gossypium hirsuturn L.） and one interspecific （cross between Gossypium hirsutum L. and Gossypium bardence L.） F2 populations were constructed by using a highly resistant cultivar and crossing it to a susceptible cultivar with 154, 79, and 148 offsprings, respectively. Simple sequence repeats （SSR） were used to screen genomic regions closely linked to FW resistance. The results showed that five QTLs associated with FW resistance were detected in two intraspecific populations using a composite interval mapping method under four different conditions. Four of these loci located on Chr. 2/Chr. 17 neighboring markers JESPR304 or CIR305 which explained 13.1 to 45.9% of the phenotypic effect. Furthermore, JESPR304 and CIR305 were previously testified and found to be tightly linked. It is possible that these four QTLs detected under different conditions were the same resistance QTL/gene. We consider that there is the possibility of a major FW resistant gene in intraspecific populations. In the interspecific mapping populations two QTLs were detected on Chr. 9 and Chr. 12/26 which explained great phenotypic variance of 49.4 and 45.7%. As the location of QTLs for FW resistance among the intraspecific and the interspecfic populations were totally different, it is suggested that there may be different resistance mechanisms between G. bardence L. and G. hursutum L. Thus, the present research provides an opportunity to understand the genetic control of resistance to FW in Gossypium hirsutum and Gossypium bardence and to conduct MAS in breeding programs to develop FW resistant cultivars.

Screening and identification of candidate Fusarium wilt-resistance genes from pumpkin

Pumpkin(Cucurbita moschata)has been widely used as cucumber(Cucumis sativus L.)rootstock to defend against Fusarium wilt(FW)and increase cucumber yields and profits.However,the resistance genes and mechanisms underlying the FW tolerance in pumpkin are poorly understood.Here we analyzed the transcriptome of pumpkin inoculated with the cucumber FW causal agent Fusarium oxysporum f.sp.cucumerinum(Foc),and obtained 3152 and 4735 upregulated genes induced by Foc at 24h after Foc inoculation compared with at 0h and 24h non-inoculated control,respectively.Next,404 common differentially expressed genes(DEGs)were screened using the criterion log_(2) FPKM(fold change)≥2.In total,206 of 404 DEGs were predominantly expressed in roots,which is the first tissue that Foc contacts and invades.140 DEGs were selected and classified into four groups(pathogenesis resistance,secondary metabolism-related,transcription factor and signal binding)based on their functional descriptions.Then,29 genes having high expression levels were selected to investigate the expression patterns induced by a Foc inoculation.Among them,16 genes were significantly induced by Foc and showed high expression levels at various treatment time points.These candidate genes may act as positive regulators of FW resistance in pumpkin and provide effective resources for improving cucumber FW resistance through breeding programs.

Efficacy evaluation and mechanism of Bacillus subtilis EBS03 against cotton Verticillium wilt

Background:In our previous study,a strain EBS03 with good biocontrol potential was screened out of 48 strains of cotton endophyte Bacillus subtilis by evaluating the controlling effect against cotton Verticillium wilt.However,its mechanism for controlling Verticillium wilt remains unclear.The objective of this study was to further clarify its con-trolling effect and mechanism against cotton Verticillium wilt.Results:The results of confrontation culture test and double buckle culture test showed that the inhibitory effects of EBS03 volatile and nonvolatile metabolite on mycelium growth of Verticillium dahliae were 70.03%and 59.00%,respectively;the inhibitory effects of sporulation and microsclerotia germination were 47.16%and 70.06%,respec-tively.In the greenhouse test,the EBS03 fermentation broth root irrigation had the highest controlling effect at 87.11%on cotton Verticillium wilt,and significantly promoted the growth of cotton seedlings.In the field experi-ment,the controlling effect of EBS03 fermentation broth to cotton Verticillium wilt was 42.54%at 60 days after cotton sowing,and the boll number per plant and boll weight in EBS03 fermentation broth seed soaking,root irrigation,and spraying treatments significantly increased by 19.48%and 7.42%,30.90%and 2.62%,15.99%and 9.20%,respec-tively.Furthermore,EBS03 improved the resistance of cotton leaves against the infection of V.dahliae,and induced the outbreak of reactive oxygen species and accumulation of callose.In addition,the results of real time fluorescent quantitative polymerase chain reaction(RT-qPCR)detection showed that EBS03 significantly induced upregulation expression level of defense-related genes PAL,POD,PPO,and PR10 in cotton leaves,enhanced cotton plant resistance to V.dahliae,and inhibited colonization level of this fungal pathogen in cotton.Conclusion:Bacillus subtilis EBS03 has a good biological defense capability,which can inhibit the growth and coloni-zation level of V.dahliae,and activate the resistance of cotton to Verticillium wilt,thus increase cotton yield.

Progresses in the Mechanism of Resistance to Fusarium Wilt in Cucumber (Cucumis sativus L.)

Fusarium wilt caused by Fusarium oxysporum f.sp. cucumerinum （Owen） is one of the most devastating diseases in cucumber production worldwide. Recent progresses in the mechanism of resistance to Fusarium wilt in cucumber were reviewed in this paper, including pathogenic mechanism of Fusarium oxysporum, the resistance mechanism of cucumber, the heredity of resistance, and the location of resistance genes. Following works should be the location and cloning of resistance genes with molecular biologic methods.

GbAt11 gene cloned from Gossypium barbadense mediates resistance to Verticillium wilt in Gossypium hirsutum

Background:Gossypium hirsutum is highly susceptible to Verticillium wilt,and once infected Verticillium wilt,its yield is greatly reduced.But G.barbadense is highly resistant to Verticillium wilt.It is possible that transferring some disease-resistant genes from G.barbadense to G.hirsutum may contribute to G.hirsutum resistance to Verticillium wilt.Result:Here,we described a new gene in G.barbadense encoding AXMN Toxin Induced Protein-11,Gb At11,which is specifically induced by Verticillium dahliae in G.barbadense and enhances Verticillium wilt resistance in G.hirsutum.Overexpression in G.hirsutum not only significantly improves resistance to Verticillium wilt,but also increases the boll number per plant.Transcriptome analysis and real-time polymerase chain reaction showed that Gb At11 overexpression can simultaneously activate FLS2,BAK1 and other genes,which are involved in ETI and PTI pathways in G.hirsutum.Conclusion:These data suggest that Gb At11 plays a very important role in resistance to Verticillium wilt in cotton.And it is significant for improving resistance to Verticillium wilt and breeding high-yield cotton cultivars.

Identification of Molecular Markers Linked to the Wilt Resistance Gene FuJ7(t) in Flax

A cross between wilt resistant flax variety Jinya7 and susceptible variety Jinya1 wasmade for mapping wilt resistance gene(s). The inoculation test of F1 and F2 progeny provedthat the resistance of Jinya7 to wilt is controlled by two dominant genes. With 48 EcoRⅠ/MseⅠ primer combinations, amplified fragment length polymorphisms (AFLP) analysis wasperformed on two parents and their F2 resistance and susceptibility bulks. A total ofabout 3300 distinguishable bands were amplified, of which three bands had stabledifferences. The genetic linkage analysis of the three polymorphic DNA fragments withthe resistance gene(s) was made in the F2 segregating population derived from the crossbetween Jinya7 and Jinya1. The DNA fragment AG/CAG was found closely linked to one of thewilt-resistant genes, which with a genetic distance of 5.2cm, was tentatively named FuJ7(t).The cloned fragment AG/CAG was sequenced and then converted successfully to a sequencecharacterized amplified region (SCAR) marker, which can be used more conveniently in theidentification and marker-assisted selection for the wilt resistance gene FuJ7(t) toflax wilt.

Utilization and Inheritance of Watermelon Resistance to Fusarium oxysporum f.sp.niveum Introduced from Bottle Gourd

Sugar Baby susceptible to Fusarium oxysporum f. sp. niveum was crossed with high resistant male parents D3 - 1 and D3 - 2, respectively. F1 hybrids showed high resistance. The segregation ratios of resistance to susceptibility of F2 and of BC1 hybrid population from Sugar Baby tallied with 3: 1 and 1: 1, respectively. The results indicated that the resistance to Fusarium wilt was a kind of dominant inheritance controlled by mono- gene or mono- segment DNA. Furthermore, 5 hybrid combinations with fine character were bred. Among them, 3 were high resistant to Fusarium wilt,and 2 were medium res

In vitro Screening of Fusarium Wilt-resistant Germplasm Resources of Red Edible Seed Watermelon( Citrullus lanatus ssp. vulgaris var. megalaspermus Lin et Chao)

[ Objective] This study aimed to establish an appropriate technology system for in vitro screening of Fuzarium wilt-resistant germplasm resources of red edible seed watermelon and obtain variants resistant to fusaric acid, thus providing resistant materials for breeding Fusarium wilt-resistant red edible seed watermel- on. [ Method] Using Zhongxin No. 1 red edible seed watermelon advemitious buds as screening materials and fusaric acid （FA） as a stress agent, in vitro screen- ing of Fusarium wilt-resistant red edible seed watermelon clonal variants and identification of Fusarium wilt-resistance of the germplasm resources of red edible seed watermelon were performed. [ Result] The results showed that the appropriate FA for in vitro screening of Fusarium wilt-resistant red edible seed watermelon vari- ants was 15 mg/L. In vitro screening system for Fusarium wilt-resistant red edible seed watermelon variants was established preliminarily and FA-resistant regenera- ted plants were obtained. Among the 36 germplasm resources of red edible seed watermelon, there were 2 highly resistant materials, 6 moderately resistant materi- als, 11 slightly resistant materials and 17 highly susceptible materials. [ Conclusion] This study confirmed preliminarily that in vitro screening method is effective for obtaining resistant materials of red edible seed watermelon.

Succinate dehydrogenase SDH1–1 positively regulates cotton resistance to Verticillium dahliae through a salicylic acid pathway

Background:Verticillium wilt,caused by the soil-borne fungus of Verticillium dahliae Kleb.,is one of the most devastating diseases of cotton.The complex mechanism underlying cotton resistance to Verticillium wilt remains uncharacterized.Identifying an endogenous resistance gene may be helpful to control this disease.Previous studies revealed that succinate dehydrogenase(SDH)is involved in reactive oxygen species(ROS)-induced stress signaling pathway that is likely to be triggered by salicylic acid(SA).Here,through the metabolomics and differential expression analyses in wilt-inoculated cotton(Gossypium hirsutum),we noticed that Gh SDH1–1 gene in cotton may play an important role in the resistance to V.dahlia.Then we reported Gh SDH1–1 gene and its functional analysis in relation to the resistance of cotton to V.dahliae.Results:The Gh SDH1–1 gene in cotton root was significantly up-regulated after V.dahlia inoculation,and its expression level peaked at 12 and 24 h post-infection.SA can also induce the up-regulation of Gh SDH1–1.Additionally,the functional analysis showed that Gh SDH1–1-silenced cotton was more susceptible to V.dahliae than the control because of the significant decrease in abundance of immune-related molecules and severe damage to the SA-signaling pathway.In Arabidopsis thaliana,high expression of Gh SDH1–1 conferred high resistance to V.dahliae.Arabidopsis that overexpressed Gh SDH1–1 had higher resistance to V.dahliae infection compared with the wild-type.Conclusions:Our findings provide new insights into the role of Gh SDH1–1;it positively regulates cotton resistance to Verticillium wilt.The regulatory mechanism of Gh SDH1–1 is closely related to SA-related signaling pathway.

基于BSA-seq技术对西瓜抗枯萎病生理小种1的基因定位

西瓜枯萎病是由尖孢镰刀菌西瓜专化型(Fusarium oxysporum f.sp.niveum)引起的土传真菌病害,是西瓜生产的限制因子。本研究以西瓜抗病种质“V13-9-3”和感病种质“W16-1”为亲本,构建了“W16-1”דV13-9-3”的F1、F2群体。苗期人工接种结果显示:F1植株表现抗病,F2群体的抗病∶感病分离比为3∶1,表明西瓜枯萎病生理小种1的抗性遗传受显性单基因控制。利用BSA技术,采用SNP指数和InDel指数法,将西瓜抗枯萎病基因Fon-1定位在1号染色体上2.20 Mb和2.32 Mb区域内,将这2种方法的定位结果取交集,最终将目标抗病基因区域缩小在2.20 Mb之内,该区域包含8个非同义突变基因和1个移码突变基因。利用荧光定量PCR检测这9个突变基因在接种枯萎病菌5 d和10 d后的表达量变化,发现抗病亲本“V13-9-3”中有7个基因的表达量显著高于感病亲本“W16-1”。研究通过结合SNP指数和InDel指数分析可以快速进行基因初定位,为西瓜抗枯萎病的分子机理研究提供理论依据。

Gh_D11G050000参与棉花黄萎病抗性的功能分析

【目的】克隆Gh_D11G050000基因,解析其在棉花黄萎病抗性反应中的功能和作用机制,为开展棉花抗病分子育种提供理论依据。【方法】利用生物信息学方法分析Gh_D11G050000基因的序列特征和系统进化关系。通过实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qRT-PCR)分析该基因的表达模式以及大丽轮枝菌(Verticillium dahliae)侵染后的表达量变化。利用病毒诱导的基因沉默(virus-induced gene silencing, VIGS)技术初步验证该基因的功能。通过转录组测序分析和相关基因的表达量测定初步解析其抗病机制。【结果】Gh_D11G050000与Gh_A11G049600和Gr_11G034620蛋白的亲缘关系最近。qRT-PCR分析发现,Gh_D11G050000在棉花根部的表达量最高;该基因的表达量在大丽轮枝菌侵染后显著增加。Gh_D11G050000沉默植株对大丽轮枝菌的抗性减弱,具体表现为茎秆维管束褐变加重,有病菌繁殖的茎段数量明显增加,病株率和病情指数显著升高。转录组及qRT-PCR分析表明,Gh_D11G050000沉默棉株中茉莉酸、乙烯信号通路和木质素合成途径中相关基因的转录水平降低。【结论】Gh_D11G050000通过影响木质素合成基因、茉莉酸和乙烯信号通路相关基因的表达正调控棉花对黄萎病的抗性。

甜瓜抗枯萎病资源筛选及抗病生理指标相关性分析

为明确黑龙江地区甜瓜枯萎病生理小种,探究枯萎病胁迫下甜瓜生理指标变化,完善甜瓜枯萎病抗病评价体系,利用表型鉴定结合分子标记辅助选择(MAS,marker-assisted selection)对24份甜瓜资源进行抗病筛选。结果表明,黑龙江省尖孢镰刀菌甜瓜专化型Fm-1为1号生理小种。分子标记辅助选择共筛选出抗病资源15份,感病资源9份,具有较高的筛选效率。其中野生型资源1314抗性最强,发病率仅为10%;栽培种22-152的感病程度最高。利用筛选的两个抗性极端材料在幼苗期接种病原菌并于根部取样,测定抗病生理指标并进行相关性分析。结果表明,枯萎病胁迫下的抗病和感病材料生理指标响应差异显著,过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性与发病率呈极显著负相关;丙二醛含量和相对电导率与发病率呈极显著正相关。综上,利用功能性分子标记可在早期实现精准筛选甜瓜抗枯萎病品种,相对电导率和CAT及PAL活性可作为抗枯萎病评价的首选指标。

苦瓜枯萎病抗性生理研究及鉴定评价体系构建

通过苦瓜枯萎病抗性生理研究,构建高效精准的苦瓜枯萎病抗性鉴定评价体系,旨在为苦瓜抗枯萎病种质资源筛选及抗病品种选育提供理论基础。以热科2号苦瓜为试验材料,选用灌根法、胚根法和伤根法在苦瓜2叶1心期接种不同浓度的苦瓜枯萎病菌,分析不同接种时间的病情指数和生理响应的差异情况,以及不同接种方法和接种浓度对病害的影响。结果表明,采用灌根法接种1×10^(6)个·mL^(-1)的枯萎病菌,在第25天时病情指数为37.1,显著低于1×10^(7)个·mL^(-1)、显著高于1×10^(4)个·mL^(-1),苯丙氨酸解氨酶活性和叶绿素含量显著低于胚根法,多酚氧化酶活性与可溶性蛋白含量与胚根法相比无显著差异。在苦瓜幼苗2叶1心时采用灌根法接种1×10^(6)个·mL^(-1)的枯萎病菌,25 d时鉴定发病情况,可作为精准高效鉴定苦瓜枯萎病抗性的体系。