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Combining simplified DNA extraction technology and recombinase polymerase amplification assay for rapid and equipment-free detection of citrus pathogen Phytophthora parasitica
CHEN Wei-yu, YU Jia, XU Heng, LU Xin-yu, DAI Ting-ting, TIAN Yue-e, SHEN Dan-yu, DOU Dao-long
2021, 20 (10): 2696-2705.   DOI: 10.1016/S2095-3119(20)63459-1
Abstract125)      PDF in ScienceDirect      
Foot and root rot caused by Phytophthora parasitica is a substantial threat to citrus cultivation, affecting both yield and quality.  Thus, rapid and accurate detection of P. parasitica plays an important role in disease management.  The aim of this study was to develop a simple diagnostic method to detect P. parasitica infection by combining recombinase polymerase amplification and lateral flow strips (LF-RPA).  To establish the LF-RPA assay of P. parasitica, the primers and probe designed based on the Ypt1 gene were tested for specificity to P. parasitica, which showed no cross-reactivity with DNAs of other related oomycete species.  The LF-RPA assay detected the amount of genomic DNA of P. parasitica which was as low as 1 pg.  To make the LF-RPA assay useful in low-resource settings, four simplified DNA extraction methods were compared, after which the LF-RPA assay was applied, with no specialized equipment, to analyze a diverse range of citrus tissues by using a simplified PEG-NaOH method for DNA extraction.  This method was successful in detecting P. parasitica in infected plant samples within 30 min.  Combining the LF-RPA assay and a simplified DNA extraction method could be a potential  detection test for P. parasitica, especially in areas with limited resources.
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Influence of Climate and Socio-Economic Factors on the Spatio-Temporal Variability of Soil Organic Matter: A Case Study of Central Heilongjiang Province, China
SHI Shu-qin, CAO Qi-wen, YAO Yan-min, TANG Hua-jun, YANG Peng, WU Wen-bin, XU Heng-zhou, LIU Jia , LI Zheng-guo
2014, 13 (7): 1486-1500.   DOI: 10.1016/S2095-3119(14)60815-7
Abstract1734)      PDF in ScienceDirect      
For the scientific management of farmland, it is significant to understand the spatio-temporal variability of soil organic matter and to study the influences of related factors. Using geostatistical theory, GIS spatial analysis, trend analysis and a Geographically Weighted Regression (GWR) model, this study analyzed the response of soil organic matter to climate and socio-economic factors in central Heilongjiang Province during the past 25 years. Second soil survey data of China for 1979-1985, 2005 field sampling data, climate observations and socio-economic data for 1980-2005 were analyzed. First, soil organic matter in 2005 was spatially interpolated using the Co-Kriging method along with auxiliary data sets of soil type and pH. The spatio-temporal variability was then studied by comparison with the 1980s second soil census data. Next, the temporal trends in climate and socio-economic factors over the past 25 years were investigated. Finally, we examined the variation of the response of soil organic matter to climate and socio-economic factors using the GWR model spatially and temporally. The model showed that 53.82% area of the organic matter content remained constant and 29.39% has decreased during the past 25 years. The impact of precipitation on organic matter content is mainly negative, with increasing absolute values of the regression coefficient. The absolute value of regression coefficient of annual average temperature has decreased, and more areas are now under its negative effects. In addition, the areas of positive regression coefficient of annual sunshine hours have northward shifted, with the increasing absolute value of positive coefficient and decreasing absolute value of negative coefficient. The areas of positive regression coefficient of mechanized farming as a socio-economic factor have westward shifted, with the increasing absolute value of negative coefficient and decreasing absolute value of positive coefficient. The area of regions with the positive regression coefficient of irrigation has expanded. The regions with positive regression coefficient of fertilizer use have shrinked. The positive regression coefficient of mulch film consumption has significantly increased. The regression coefficient of pesticide consumption was mainly positive in the west of the study area, while it was negative to the east. Generally, GWR model is capable to investigate the influence of both climatic and socio-economic factors, avoided the insufficiency of other research based on the single perspective of climatic or socio-economic factors. Therefore, we can conclude that GWR model could provide methodological support for global change research and serve as basic reference for cultivated land quality improvement and agricultural decision making.
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Transgenic Expression of a Functional Fragment of Harpin Protein Hpa1 in Wheat Represses English Grain Aphid Infestation
XU Man-yu, ZHOU Ting, ZHAO Yan-ying, LI Jia-bao, XU Heng, DONG Han-song , ZHANG Chun-ling
2014, 13 (12): 2565-2576.   DOI: 10.1016/S2095-3119(13)60735-2
Abstract1451)      PDF in ScienceDirect      
The harpin protein Hpa1 produced by the rice bacterial blight pathogen promotes plant growth and induces plant resistance to pathogens and insect pests. The region of 10-42 residues (Hpa110-42) in the Hpa1 sequence is critical as the isolated Hpa110-42 fragment is 1.3-7.5-fold more effective than the full length in inducing plant growth and resistance. Here we report that transgenic expression of Hpa110-42 in wheat induces resistance to English grain aphid, a dominant species of wheat aphids. Hpa110-42-induced resistance is effective to inhibit the aphid behavior in plant preference at the initial colonization stage and repress aphid performances in the reproduction, nymph growth, and instar development on transgenic plants. The resistance characters are correlated with enhanced expression of defense-regulatory genes (EIN2, PP2-A, and GSL10) and consistent with induced expression of defense response genes (Hel, PDF1.2, PR-1b, and PR-2b). As a result, aphid infestations are alleviated in transgenic plants. The level of Hpa110-42-induced resistance in regard to repression of aphid infestations is equivalent to the effect of chemical control provided by an insecticide. These results suggested that the defensive role of Hpa110-42 can be integrated into breeding germplasm of the agriculturally significant crop with a great potential of the agricultural application.
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