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Molecular diagnosis and direct quantification of cereal cyst nematode (Heterodera filipjevi) from field soil using TaqMan real-time PCR
JIAN Jin-zhuo, HUANG Wen-kun, KONG Ling-an, JIAN Heng, Sulaiman ABDULSALAM, PENG De-liang, PENG Huan
2023, 22 (8): 2591-2601.   DOI: 10.1016/j.jia.2022.09.016
Abstract221)      PDF in ScienceDirect      
Heterodera filipjevi continues to be a major threat to wheat production worldwide. Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease. In the present study, a TaqManminor groove binder (TaqMan-MGB) probe-based fluorescence quantitative real-time PCR (qPCR) was successfully developed and used for quantifying H. filipjevi from DNA extracts of soil. The primers and probe designed from the obtained RAPD-SCAR marker fragments of H. filipjevi showed high specificity to H. filipjevi using DNA from isolatesconfirmed species of 23 Heterodera spp., 1 Globodera spp. and 3 Pratylenchus spp. The qPCR assay is highly sensitive and provides improved H. filipjevi detection sensitivity of as low as 4–3 single second-stage juvenile (J2) DNAs, 10–3 female DNAs, and 0.01 μg μL–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 H. filipjevi in naturally infested field soils. There was a high correlation between the H. filipjevi numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR assay. qPCR potentially provides a useful platform for the efficient detection and quantification of H. filipjevi directly from field soils and to quantify this species directly from DNA extracts of field soils
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Potassium sulphate induces resistance of rice against the root-knot nematode Meloidogyne graminicola
LIU Mao-Yan, PENG De-liang, SU Wen, XIANG Chao, JIAN Jin-zhuo, ZHAO Jie, PENG Huan, LIU Shi-ming, KONG Ling-an, DAI Liang-ying, HUANG Wen-kun, LIU Jing
2022, 21 (11): 3263-3277.   DOI: 10.1016/j.jia.2022.08.002
Abstract268)      PDF in ScienceDirect      

Potassium (K), an important nutrient element, can improve the stress resistance/tolerance of crops.  The application of K in resisting plant-parasitic nematodes shows that the K treatment can reduce the occurrence of nematode diseases and increase crop yield.  However, data on K2SO4 induced rice resistance against the root-knot nematode Meloidogyne graminicola are still lacking.  In this work, K2SO4 treatment reduced galls and nematodes in rice plants and delayed the development of nematodes.  Rather than affecting the attractiveness of roots to nematodes and the morphological phenotype of giant cells at feeding sites, such an effect is achieved by rapidly priming hydrogen peroxide (H2O2) accumulation and increasing callose deposition.  Meanwhile, galls and nematodes in rice roots were more in the potassium channel OsAKT1 and transporter OsHAK5 gene-deficient plants than in wild-type, while the K2SO4-induced resistance showed weaker in the defective plants.  In addition, during the process of nematode infection, the expression of jasmonic acid (JA)/ethylene (ET)/brassinolide (BR) signaling pathway-related genes and pathogenesis-related (PR) genes OsPR1a/OsPR1b was up-regulated in rice after K2SO4 treatment.  In conclusion, K2SO4 induced rice resistance against M. graminicola.  The mechanism of inducing resistance was to prime the basal defense and required the participation of the K+ channel and transporter in rice.  These laid a foundation for further study on the mechanism of rice defense against nematodes and the rational use of potassium fertilizer on improving rice resistance against nematodes in the field.

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A fragment of a 70-kDa Heterodera glycines heat shock protein (HgHSP70) interacts with soybean cyst nematode-resistant protein GmSHMT08
LIU Zhi, ZHANG Liu-ping, ZHAO Jie, JIAN Jin-zhuo, PENG Huan, HUANG Wen-kun, KONG Ling-an, PENG De-liang, LIU Shi-ming
2022, 21 (10): 2973-2983.   DOI: 10.1016/j.jia.2022.07.048
Abstract228)      PDF in ScienceDirect      


Soybean cyst nematode (SCN) Heterodera glycines is considered as the major constraint to soybean production.  GmSHMT08 at Rhg4 locus on chromosome 08, encoding a serine hydroxylmethyltransferase, is a major gene underlying resistance against Hglycines in Peking-type soybeans.  However, the molecular mechanism underpinning this resistance is less well characterized, and whether GmSHMT08 could interact with proteins in Hglycines remains unclear.  In this study, yeast two-hybrid screening was conducted using GmSHMT08 as a bait protein, and a fragment of a 70-kDa heat shock protein (HgHSP70) was screened from Hglycines that exhibited interaction with GmSHMT08.  This interaction was verified by both GST pull-down and bimolecular fluorescence complementation assays.  Our finding reveals HgHSP70 could be applied as a potential candidate gene for further exploring the mechanism on GmSHMT08-mediated resistance against SCN Hglycines.


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