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Assaying the potential of twenty-one legume plants in Medicago truncatula and M. sativa for candidate model plants for investigation the interactions with Heterodera glycines

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Journal of Integrative Agriculture 2016年第3期15卷 702-704页

作者： KONG Ling-an WU Du-qing huang wen-kun PENG Huan HE wen-ting PENG De-liang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Soybean cyst nematode Heterodera glycines is one of the most serious soil-borne pathogens in soybean production. However, the researches were limited in China due to lack of an effective pathosystem. In this study, we screened 21 legume Medicago plants in both Medicago truncatula and Medicago sativa to obtain candidate model plants for establishing a new pathosystem for legume-H. glycines interactions. The nematode infection of tested plants was assayed with Race 3 and 4 respectively, which were two dominant H. glycines inbred races in China soybean producing areas. The results showed that the model legume plant M. truncatula A17 failed to allow Race 3 of H. glycines to complete its life cycle, in contrast, it provided the Race 4 population to form several cyst nematodes, however, the female index（FI） value was approximately 1.6. Three M. sativa cultivars, including Xunlu, Aergangjin and Junren, provided either Race 3 or 4 of H. glycines to develop into mature cysts with their FI value below 5 as well. Our results demonstrated that legume plants in both M. truncatula and M. sativa were not likely to be a model plant for H. glycines because of an extreme high resistance.

关键词： soybean cyst nematode alfalfa life cycle female index pathosystem

Molecular and morphological characterization of stunt nematodes of wheat, maize, and rice in the savannahs of northern Nigeria

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Journal of Integrative Agriculture 2022年第2期21卷 586-595页

作者： Sulaiman ABDULSALAM PENG Huan LIU Shi-ming huang wen-kun KONG Ling-an PENG De-liang State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection Chinese Academy of Agricultural SciencesBeijing 100193P.R.China Division of Agricultural Colleges Department of Crop ProtectionAhmadu Bello UniversityZaria 810107Nigeria

Stunt nematodes(Tylenchorhynchus spp.)are obligate migratory root ecto-parasitic nematodes found in the fields of many cultivated *** nematodes,with phyto-sanitary potential,are frequently ignored or misdiagnosed as pests,and this may pose a threat to food *** accuracy of its identification based on a morphological approach has been challenged recently,due to the overlapping of the morphological and morphometric characters of the ***,the objective of this study is to identify and characterize stunt nematodes present in 54 fields cultivated with cereal crops(wheat,maize and rice)in the savannahs of northern Nigeria,using integrative taxonomy and molecular *** molecular and morphological studies identified and confirmed the presence of *** as the occurring specie in the savannahs of northern *** phylogenetic analysis was carried out using the internal transcribed spacer(ITS)and 28S genes of ribosomal DNA further confirmed the presence of *** first molecular characterization and sequences of the ITS and 28S rDNA gene for *** from Nigeria were provided by this ***,according to our literature search,this is the first report on *** in wheat,maize and rice in the savannahs of northern *** study to test the pathogenicity of the parasitic nematode species found in this survey is recommended for the prioritization and development of efficient management strategies.

关键词： plant-parasitic nematodes food security Tylenchorhynchus annulatus ITS phylogeny

Effect of Aspergillus niger NBC001 on the soybean rhizosphere microbial community in a soybean cyst nematode-infested field

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Journal of Integrative Agriculture 2021年第12期20卷 3230-3239页

作者： JIN Na LIU Shi-ming PENG Huan huang wen-kun KONG Ling-an PENG De-liang Department of Horticulture Beijing Vocational College of AgricultureBeijing 102442P.R.China State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China

Soybean cyst nematode(SCN,Heterodera glycines Ichinohe)is one of the most important pests causing considerable damage to soybean(Glycine max(L.)Merr.)around the *** provides a strategy for sustainable nematode ***,Aspergillus niger NBC001 was isolated from the cysts Heterodera *** able to control *** and promote the growth of soybean in a pot *** this study,the effects of NBC001 on *** density and on the soybean rhizosphere microbial community in a soybean cyst nematode-infested field were *** results showed that NBC001 could suppress *** by 31.7%in the ***-throughput sequencing analysis showed that NBC001 had no significant influence on soybean rhizosphere microbial community structure,indicating that seed coatdressing with the concentrated culture filtrate of NBC001 was safe for the soil ecological *** addition,high-throughput sequencing results demonstrated that at 10 days post transplantation,NBC001 increased the abundances of Actinobacteria and Acidobacteria,but decreased the abundances of Bacteroidetes and Gemmatimonadetes at the phylum ***,the abundances of Phyllobacterium,Ralstonia and H16 were increased,while the abundances of Adhaeribacter,Gemmatimonas,Sphingomonas,Flavisolibacter\Nere suppressed by application of ***,at 90 days post transplantation,NBC001 only increased the abundances oi Aeromicrobium and RB41 whereas it decreased the abundance of *** results indicated that application of NBC001 increased the relative abundances of the beneficial microorganisms such as Actinobacteria,Acidobacteria,Aeromicrobium and Phyllobacterium in the *** summary,NBC001 is an eco-friendly biocontrol agent for *** control.

关键词： Aspergillus niger NBC001 biocontrol Heterodera glycines high-throughput sequencing microbial community

Molecular diagnosis and direct quantification of cereal cyst nematode(Heterodera filipjevi) from field soil using TaqMan real-time PCR

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Journal of Integrative Agriculture 2023年第8期22卷 2591-2601页

作者： JIAN Jin-zhuo huang wen-kun KONG Ling-an JIAN Heng Sulaiman ABDULSALAM PENG De-liang PENG Huan State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China Key Laboratory of Plant Pathology of Ministry of Agriculture and Rural Affairs College of Plant ProtectionChina Agricultural UniversityBeijing100193P.R.China Division of Agricultural Colleges/Department of Crop Protection Ahmadu Bello UniversityZaria 810107Nigeria

Heterodera filipjevi continues to be a major threat to wheat production *** detection and quantification of cyst nematodes are essential for more effective control against this nematode *** the present study,a TaqManminor groove binder(TaqMan-MGB)probe-based fluorescence quantitative real-time PCR(qPCR)was successfully developed and used for quantifying *** from DNA extracts of *** primers and probe designed from the obtained RAPD-SCAR marker fragments of *** showed high specificity to *** using DNA from isolatesconfirmed species of 23 Heterodera spp.,1 Globodera *** 3 Pratylenchus *** qPCR assay is highly sensitive and provides improved *** detection sensitivity of as low as 4^(-3) single second-stage juvenile(J2)DNAs,10^(-3) female DNAs,and 0.01μgμL^(-1) genomic DNAs.A standard curve relating to the threshold cycle and log values of nematode numbers was generated and validated from artificially infested soils and was used to quantify *** in naturally infested field *** was a high correlation between the *** numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR *** potentially provides a useful platform for the efficient detection and quantification of *** directly from field soils and to quantify this species directly from DNA extracts of field soils.

关键词： cereal cyst nematode Heterodera filipjevi molecular diagnosis quantification TaqMan real-time PCR

Chemical mutagenesis and soybean mutants potential for identification of novel genes conferring resistance to soybean cyst nematode

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Journal of Integrative Agriculture 2018年第12期17卷 2734-2744页

作者： GE Feng-yong ZHENG Na ZHANG Liu-ping huang wen-kun PENG De-liang LIU Shi-ming Institute of Plant Protection Chinese Academy of Agricultural Sciences College of Plant Protection Hunan Agricultural University

The resistance of soybean(Glycine max(L.) Merr.) to soybean cyst nematode(SCN, Heterodera glycines Ichinohe), which is a devastating pathogen in soybean production and causes a large quantity of annual yield loss worldwide, can shift during the long-term interaction and domestication. It is vital to identify more new resistance genetic sources for identification of novel genes underlying resistance to SCN for management of this pathogen. In the present study, first, two ethane methylsulfonate-mutagenesis soybean M2 populations of PI 437654, which shows a broad resistance to almost all of SCN races, and Zhonghuang 13, which is a soybean cultivar in China conferring strong resistance to lodging, were developed. Many types of morphological phenotypes such as four-and five-leaflet leaves were observed from these two soybean M2 populations. Second, 13 mutants were identified and confirmed to exhibit alteration of resistance to SCN race 4 through the forward genetic screening of 400 mutants of the PI 437654 M2 population, the rate of mutants with alteration of SCNinfection phenotype is 3.25%. Third, these identified mutants were further verified not to show any changes in the genomic sequences of the three known SCN-resistant genes, GmSHMT08, GmSNAP18 and GmSANP11, compared to the wildtype soybean; and all of them were still resistant to SCN race 3 similar to the wild-type soybean. Taken together, we can conclude that the 13 mutants identified in the present study carry the mutations of the new gene(s) which contribute(s) to the resistance to SCN race 4 in PI 437654 and can be potentially used as the genetic soybean sources to further identify the novel SCN-resistant gene(s).

关键词： soybean ethane methylsulfonate-mutagenesis populations mutants soybean cyst nematode race 4 resistance

Molecular characterization and functional analysis of two new lysozyme genes from soybean cyst nematode(Heterodera glycines)

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Journal of Integrative Agriculture 2019年第12期18卷 2806-2813页

作者： WANG Ning PENG Huan LIU Shi-ming huang wen-kun Ricardo Holgado Jihong Liu-Clarke PENG De-liang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China NIBIO Norwegian Institute of Bioeconomy ResearchPb 115N0-1431 AsNorway

Soybean cyst nematode(SCN, Heterodera glycines(I.)) is one of the most important soil-borne pathogens for soybeans. In plant parasitic nematodes, including SCN, lysozyme plays important roles in the innate defense system. In this study, two new lysozyme genes(Hg-lys1 and Hg-lys2) from SCN were cloned and characterized. The in situ hybridization analyses indicated that the transcripts of both Hg-lys1 and Hg-lys2 accumulated in the intestine of SCN. The q RT-PCR analyses showed that both Hg-lys1 and Hg-lys2 were upregulated after SCN second stage juveniles(J2 s) were exposed to the Grampositive bacteria Bacillus thuringiensis, Bacillus subtilis or Staphylococcus aureus. Knockdown of the identified lysozyme genes by in vitro RNA interference caused a significant decrease in the survival rate of SCN. All of the obtained results indicate that lysozyme is very important in the defense system and survival of SCN.

关键词： soybean cyst nematode lysozyme Hg-lys1 Hg-lys2 innate defense against bacteria

Conidia of one Fusarium solani isolate from a soybean-production field enable to be virulent to soybean and make soybean seedlings wilted

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Journal of Integrative Agriculture 2018年第9期17卷 2042-2053页

作者： ZHENG Na ZHANG Liu-ping GE Feng-yong huang wen-kun KONG Ling-an PENG De-liang LIU Shi-ming Institute of Plant Protection Chinese Academy of Agricultural Sciences College of Plant Protection Hunan Agricultural University

Fusarium is usually thought to cause soybean root rot, which results in a large quantity of annual yield loss in soybean production, by its secretions including Fusarium toxins and cell wall degrading enzymes, but not by the conidia themselves that do not underlie any virulence so far. Here we report that the conidia of one Fusarium solani isolate are able to be virulent to soybean and make soybean seedlings wilted alone. We isolated them from the wilted plants in a soybean-production field and molecularly identified 17 Fusarium isolates through phylogenetic analysis. Of them, except for one isolate that showed diversity of virulence to different soybeans （virulent to one soybean whereas avirulent to another soybean）, the others were all virulent to the two tested soybeans： both conidia cultures and secretions could make soybean seedlings wilted at 5 days post infection, and their virulence had dosage effects that only conidia cultures of at least 5×10^6 conidia mL-1 could show virulence to soybean; however, the sole conidia of the F. solani isolate #4 also exhibited virulence to soybean and could make soybean seedlings wilted. Finally, we developed the specific cleaved amplified polymorphic sequences （CAPS） markers to easily differentiate Fusarium isolates. The isolate #4 in this work will likely be used to investigate the new mechanism of virulence of Fusarium to soybean.

关键词： Fusarium soybean root rot conidia secretions virulence cleaved amplified polymorphic sequences (CAPS) marker

A new pathotype characterization of Daxing and huangyuan populations of cereal cyst nematode(Heterodera avenae) in China

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Journal of Integrative Agriculture 2015年第4期14卷 724-731页

作者： CUI Jiang-kuan huang wen-kun PENG Huan LIU Shi-ming WANG Gao-feng KONG Lin-an PENG De-liang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural Sciences

The cereal cyst nematode（CCN, Heteroder aavenae） causes serious yield loss on cereal crops, especially wheat, worldwide. Daxing population in Beijing City and huangyuan population in Qinghai Province, China, are two CCN populations. In this study, the CCN pathotypes of Daxing and huangyuan populations were characterized by tests on 23 standard ＂International Test Assortment＂ with the local species wenmai 19 as the susceptible control. Tested materials were grouped by three nematode populations＇ virulence on resistant genes（Rha1, Rha2, Rha3, Cre1） and nonresistant genes, varieties and lines. Both Daxing and huangyuan populations were avirulent to Ortolan（Ha1）. Barley cvs. Ortolan, Siri, Morocco, Bajo Aragon 1-1, and Martin 403-2 were all resistant to both populations. Cultivars Herta, Harlan 43 and wheat Iskamish-K-2-light were all susceptible to huangyuan population, all of them, however, were resistant to Daxing population. The other five oats were all resistant to the two tested CCN populations. Except Iskamisch K-2-light, all the other wheat cultivars（Capa, Loros×Koga, AUS 10894, and Psathias） were susceptible to Daxing population. Because the pathotypes of the two tested CCN populations in Beijing and Qinghai were not identical to any of the 13 pathotypes previously characterized by the test assortment, we classified Daxing and huangyuan populations as the new pathotypes, named Ha91.

关键词： cereal cyst nematode Heterodera avenae Daxing population huangyuan population pathotype

The effects of Trichoderma on preventing cucumber fusarium wilt and regulating cucumber physiology

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Journal of Integrative Agriculture 2019年第3期18卷 607-617页

作者： LI Mei MA Guang-shu LIAN Hua SU Xiao-lin TIAN Ying huang wen-kun MEI Jie JIANG Xi-liang Institute of Plant Protection Chinese Academy of Agricultural Sciences College of Agronomy Heilongjiang Bayi Agricultural University

In our previous studies, we identified 3 Trichoderma strains with anti-Fusarium oxysporum activity, including T. asperellum 525, T. harzianum 610, and T. pseudokoningii 886. Here, we evaluated the effects of these 3 Trichoderma strains on preventing cucumber fusarium wilt through pot culture and greenhouse culture experiments. All 3 Trichoderma strains demonstrated higher control effects toward cucumber fusarium wilt than previous studies, with efficacies over 78%. Additionally, inoculation with the 3 Trichoderma strains significantly promoted the quality and yield of cucumbers. Among the 3 strains, Trichoderma 866 was the most effective, with disease control efficacy of 78.64% and a cucumber yield increase of 33%. Furthermore, seedlings inoculated with Trichoderma exhibited significantly increased measures of plant height, stem diameter, leaf area, aboveground fresh weight, underground fresh weight, chlorophyll content, and nitric nitrogen content, as well as the activities of several stress-resistance enzymes, including superoxide dismutase(SOD), peroxidase(POD), catalase(CAT), polyphenol oxidase(PPO), and ascorbate oxidase(AAO). In addition, the plants inoculated with Trichoderma showed decreased cell membrane permeability and malondialdehyde(MDA) content in the leaves. Together, our results suggest that T. asperellum 525, T. harzianum 610, and T. pseudokoningii 886 inoculations inhibit F. oxysporum infection, stimulate the metabolism in cucumbers, and enhance the activities of stress-resistance enzymes, which consequently promote the growth of cucumber plants, prevent cucumber fusarium wilt, and improve the yield and quality of cucumbers. T. harzianum is a commonly used biocontrol fungus, while few studies have focused on T. asperellum or T. koningense. In this study, strains of T. asperellum and T. pseudokoningii showed excellent plant disease prevention and growth promoting effects on cucumber, indicating that they also have great potential as biocontrol fungi

关键词： Trichoderma cucumber fusarium wilt physicochemical features control effect Fusarium oxysporum f. sp. cucumerinum Owen