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Gene cloning,protein expression,and enzymatic characterization of a double-stranded RNA degrading enzyme in Apolygus lucorum

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Insect Science 2024年第1期31卷 119-133页

作者： Jie-Yu Zhang Jing Zhao Keyan Zhu-Salzman Qin-Qin Ji Yi-Ping Jiang Liu-Bin Xiao De-Jin Xu Guang-Chun Xu Lin-Quan Ge Yong-An Tan College of plant protection Yangzhou UniversityYangzhouJiangsu ProvinceChina Institute of plant protection Jiangsu Academy of Agricultural SciencesNanjingJiangsu ProvinceChina Department of Entomology Texas A&M UniversityCollege StationTXUSA Taizhou Customs of the People's Republic of China TaizhouJiangsu ProvinceChina

RNA interference(RNAi)is a powerful tool that post-transcriptionally silences target genes in eukaryotic cells.However,silencing efficacy varies greatly among different insect species.Recently,we met with little success when attempting to knock down genes in the mirid bug Apolygus lucorum via dsRNA injection.The disappearance of double-stranded RNA(dsRNA)could be a potential factor that restricts RNAi efficiency.Here,we found that dsRNA can be degraded in midgut fluids,and a dsRNase of A.lucorum(AldsRNase)was identified and characterized.Sequence alignment indicated that its 6 key amino acid residues and the Mg2+-binding site were similar to those of other insects’dsRNases.The signal peptide and endonuclease non-specific domain shared high sequence identity with the brown-winged green stinkbug Plautia stali dsRNase.AldsRNase showed high salivary gland and midgut expression and was continuously expressed through the whole life cycle,with peaks at the 4th instar ecdysis in the whole body.The purified AldsRNase protein obtained by heterologously expressed can rapidly degrade dsRNA.When comparing the substrate specificity of AldsRNase,3 specific substrates(dsRNA,small interfering RNA,and dsDNA)were all degraded,and the most efficient degradation is dsRNA.Subsequently,immunofluorescence revealed that AldsRNase was expressed in the cytoplasm of midgut cells.Through cloning and functional study of AldsRNase,the enzyme activity and substrate specificity of the recombinant protein,as well as the subcellular localization of nuclease,the reason for the disappearance of dsRNA was explained,which was useful in improving RNAi efficiency in A.lucorum and related species.

关键词： Apolygus lucorum cloning dsRNase enzymatic activity protein expression

AcWRKY28 mediated activation of AcCPK genes confers salt tolerance in pineapple(Ananas comosus)

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Horticultural plant Journal 2024年第2期10卷 398-412页

作者： Qiao Zhou Samaranayaka Vidana Gamage Nirosha Priyadarshani Rongjuan Qin Han Cheng Tiantian Luo Myat Hnin Wai Mohammad Aqa Mohammadi Yanhui Liu Chang Liu Hanyang Cai Xiaomei Wang Yeqiang Liu Yuan Qin Lulu Wang Guangxi Key Lab of Sugarcane Biology State Key Laboratory for Conservation and Utilization of Subtropical Agro-BioresourcesCollege of AgricultureGuangxi UniversityNanningGuangxi 530004China State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops College of Life ScienceCollege of Plant ProtectionCollege of AgricultureCollege of HorticultureFujian Provincial Key Laboratory of Haixia Applied Plant Systems BiologyCenter for Genomics and BiotechnologyFujian Agriculture and Forestry UniversityFuzhouFujian 350002China Pingtan Science and Technology Research Institute of Fujian agriculture and Forestry University PingtanFujian 350400China National Education Commission BMICHColombo00700Sri Lanka Fishery Multiplication Management station of Lijiang River Water Supply Hub Project GuilinGuangxi 541001China Horticulture Research Institute Guangxi Academy of Agricultural SciencesNanning Investigation Station of South Subtropical Fruit TreesMinistry of AgricultureNanningGuangxi 530007China

Unfavorable environmental cues severely affect crop productivity resulting in significant economic losses to farmers. In plants, multiple regulatory genes, such as the WRKY transcription factor (TF) family, modulate the expression of defense genes. However, the role of the pineapple WRKY genes is poorly understood. Here, we studied the pineapple WRKY gene, AcWRKY28, by generating AcWRKY28 over-expressing transgenic pineapple plants. Overexpression of AcWRKY28 enhanced the salt stress resistance in transgenic pineapple lines. Comparative transcriptome analysis of transgenic and wild-type pineapple plants showed that “plant-pathogen interaction” pathway genes, including 9calcium-dependent protein kinases (CPKs), were up-regulated in AcWRKY28 over-expressing plants. Furthermore, chromatin immunoprecipitation and yeast one-hybrid assays revealed AcCPK12, AcCPK3, AcCPK8, AcCPK1, and AcCPK15 as direct targets of AcWRKY28. Consistently, the study of AcCPK12 over-expressing Arabidopsis lines showed that AcCPK12 enhances salt, drought, and disease resistance. This study shows that AcWRKY28 plays a crucial role in promoting salt stress resistance by activating the expression of AcCPK genes.

关键词： Pineapple AcWRKY AcCPK Transcription factor Salt stress ChIP

PIF4 interacts with ABI4 to serve as a transcriptional activator complex to promote seed dormancy by enhancing ABA biosynthesis and signaling

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Journal of Integrative plant Biology 2024年第5期66卷 909-927页

作者： Xiaofeng Luo Yujia Dai Baoshan Xian Jiahui Xu Ranran Zhang Muhammad Saad Rehmani Chuan Zheng Xiaoting Zhao Kaitao Mao Xiaotong Ren Shaowei Wei Lei Wang Juan He Weiming Tan Junbo Du Weiguo Liu Shu Yuan Kai Shu Shaanxi Key Laboratory of Qinling Ecological Intelligent Monitoring and protection School of Ecology and EnvironmentNorthwestern Polytechnical UniversityXi'an 710129China Research&Development Institute of Northwestern Polytechnical University in ShenzhenShenzhen 518057China College of Agronomy and Biotechnology China Agricultural UniversityBeijing 100193China Institute of Ecological agriculture Sichuan Agricultural UniversityChengdu 611130China College of Resources Sichuan Agricultural UniversityChengdu 611130China

Transcriptional regulation plays a key role in the control of seed dormancy,and many transcription factors(TFs)have been documented.However,the mechanisms underlying the interactions between different TFs within a transcriptional complex regulating seed dormancy remain largely unknown.Here,we showed that TF PHYTOCHROME-INTERACTING FACTOR4(PIF4)physically interacted with the abscisic acid(ABA)signaling responsive TF ABSCISIC ACID INSENSITIVE4(ABI4)to act as a transcriptional complex to promote ABA biosynthesis and signaling,finally deepening primary seed dormancy.Both pif4 and abi4 single mutants exhibited a decreased primary seed dormancy phenotype,with a synergistic effect in the pif4/abi4 double mutant.PIF4 binds to ABI4 to form a heterodimer,and ABI4 stabilizes PIF4 at the protein level,whereas PIF4 does not affect the protein stabilization of ABI4.Subsequently,both TFs independently and synergistically promoted the expression of ABI4 and NCED6,a key gene for ABA anabolism.The genetic evidence is also consistent with the phenotypic,physiological and biochemical analysis results.Altogether,this study revealed a transcriptional regulatory cascade in which the PIF4–ABI4 transcriptional activator complex synergistically enhanced seed dormancy by facilitating ABA biosynthesis and signaling.

关键词： ABA ABI4 PIF4 seed dormancy and germination transcriptional complex

Yield Characteristics of a New High-quality Disease-resistant Wheat Variety Chuanmai 618

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plant Diseases and Pests 2024年第2期15卷 22-26,31页

作者： Jianmin ZHENG Zongjun PU Jijuan LU Jiangtao LUO Qingyan DENG Peixun LIU Shizhao LI Crop Research Institute Sichuan Academy of Agricultural Sciences/Environment-Friendly Crop Germplasm Innovation and Genetic Improvement Key Laboratory of Sichuan Province/Key Laboratory of Wheat Biology and Genetic Improvement on Southwestern ChinaMinistry of Agriculture and Rural AffairsChengdu 610066China sichuan Seed Management station Chengdu 610041China

[Objectives]The paper was to analyze the yield characteristics of a new high-quality disease-resistant wheat variety Chuanmai 618.[Methods]The yield characteristics of Chuanmai 618 were analyzed using the AMMI model and GGE biplot based on data from a 2-year regional test and a 1-year production test.[Results]The analysis of the AMMI model for the 2-year regional test indicated that Chuanmai 618 had a moderate yield and good stability.During the production test,Chuanmai 618 had an average yield of 450.52 kg/666.7 m^(2),an effective spike of 235700 spike/666.7 m^(2),a 1000-seed weight of 47.93 g,and a kernel number per spike of 47.28.The AMMI analysis sequencing graph showed that the varieties were ranked in the following order:Zhongkemai 1816>Chuanmai 618>Shumai 1958>Chuanyu 42>Mianmai 367>Xikemai 5518.According to the GE analysis,Chuanmai 618 had comparative advantages.[Conclusions]The new wheat variety Chuanmai 618 is a high-quality disease-resistant variety with good yield and stability.

关键词： Chuanmai 618 High quality Disease-resistant AMMI model High yield Stability

Fine mapping and characterization of stripe rust resistance gene YrAYH in near-isogenic lines derived from a cross involving wheat landrace Anyuehong

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The Crop Journal 2024年第3期12卷 826-835页

作者： Li Long Jue Li Linyu Huang Huiling Jin Fangnian Guan Haipeng Zhang Sasa Zhao Hao Li Zhien Pu Wei Li Qiantao Jiang Yuming Wei Jian Ma Houyang Kang Shoufen Dai Pengfei Qi Qiang Xu Mei Deng Youliang Zheng Yunfeng Jiang Matthew James Moscou Guoyue Chen State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China Sichuan Agricultural UniversityChengdu 611130SichuanChina Triticeae Research Institute Sichuan Agricultural UniversityChengdu 611130SichuanChina College of Resources Sichuan Agricultural UniversityChengdu 611130SichuanChina The Sainsbury Laboratory Norwich Research ParkNorwich NR47UHEnglandUK United States Department of agriculture-Agricultural Research Service Cereal Disease LaboratorySt.PaulMN 55108USA

Stripe rust,caused by Puccinia striiformis f.***.tritici(Pst),is a devastating disease in wheat worldwide.Discovering and characterizing new resistance genes/QTL is crucial for wheat breeding programs.In this study,we fine-mapped and characterized a stripe rust resistance gene,YRAYH,on chromosome arm 5BL in the Chinese wheat landrace Anyuehong(AYH).Evaluations of stripe rust response to prevalent Chinese Pst races in near-isogenic lines derived from a cross of Anyuehong and Taichung 29 showed that YrAYH conferred a high level of resistance at all growth stages.Fine mapping using a large segregating population of 9748 plants,narrowed the YRAYH locus to a 3.7 Mb interval on chromosome arm 5BL that included 61 annotated genes.Transcriptome analysis of two NIL pairs identified 64 upregulated differentially expressed genes(DEGs)in the resistant NILs(NILs-R).Annotations indicated that many of these genes have roles in plant disease resistance pathways.Through a combined approach of fine-mapping and transcriptome sequencing,we identified a serine/threonine-protein kinase SRPK as a candidate gene underlying YrAYH.A unique 25 bp insertion was identified in the NILs-R compared to the NILs-S and previously published wheat genomes.An InDel marker was developed and co-segregated with YrAYH.Agronomic trait evaluation of the NILs suggested that YrAYH not only reduces the impact of stripe rust but was also associated with a gene that increases plant height and spike length.

关键词： Candidate gene analysis Crop protection Puccinia striiformis Transcriptome analyses

Different responses of soil fauna gut and plant rhizosphere microbiomes to manure applications

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Soil Ecology Letters 2024年第2期6卷 57-69页

作者： Shuai Du Xiang Zhao Yue Zhang Changlong Shu Ju-Pei Shen Key Laboratory of Urban Environment and Health Ningbo Urban Environment Observation and Research StationInstitute of Urban EnvironmentChinese Academy of SciencesXiamen361021China School of Geographical Sciences Fujian Normal UniversityFuzhou350007China Institute of plant Nutrition Resources and EnvironmentBeijing Academy of Agriculture and Forestry SciencesBeijing100097China State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing100193China Hebei Key Laboratory of Soil Entomology Cangzhou Academy of Agricultural and Forestry SciencesCangzhou061001China

Microbial attributes were compared between soil fauna gut and plant rhizosphere.•Manure applications decreased or increased gut or rhizosphere bacterial diversity.•Stochastic or deterministic processes drove gut or rhizosphere bacterial assembly.•Manure applications increased bacterial network complexity of gut and rhizosphere.Diverse microbes inhabit animals and plants,helping their hosts perform multiple functions in agricultural ecosystems.However,the responses of soil fauna gut and plant rhizosphere microbiomes to livestock manure applications are still not well understood.Here we fed Protaetia brevitarsis larvae(PBL)with chicken manure and collected their frass.The frass and manure were applied as fertilizers to lettuce pots.We then compared the changes of microbial diversity,community assembly,and potential functions between the gut group(i.e.,all PBL gut and frass samples)and the rhizosphere group(i.e.,all rhizosphere soil samples).We revealed that manure applications(i.e.,feeding or fertilization)decreased bacterial diversity in the gut group but increased that in the rhizosphere group.Particularly,the proportions of Bacilli in the gut group and Gammaproteobacteria in the rhizosphere group were increased(up to a maximum of 33.8%and 20.4%,respectively)after manure applications.Stochastic and deterministic processes dominated community assembly in the gut and rhizosphere microbiomes,respectively.Manure applications increased the microbial co-occurrence network complexity of both the gut and rhizosphere groups.Moreover,the proportions of functional taxa associated with human/animal pathogens in the gut group and carbon/nitrogen cycling in the rhizosphere group were enhanced(up to 2.6-fold and 24.6-fold,respectively).Our findings illustrate the different responses of microbial diversity,community assembly,and potential functions in soil fauna gut and plant rhizosphere to manure applications.The results could enhance our knowledge on the reasonable utilization of animal and plant microbiomes in agricultural ma

关键词： soil fauna gut plant rhizosphere manure applications community assembly microbial functions

Assessment of soil total phosphorus storage in a complex topography along China's southeast coast based on multiple mapping scales

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Pedosphere 2024年第1期34卷 236-251页

作者： Zhongxing CHEN Jing LI Kai HUANG Miaomiao WEN Qianlai ZHUANG Licheng LIU Peng ZHU Zhenong JIN Shihe XING Liming ZHANG Fujian Provincial Key Laboratory of Soil Environmental Health and Regulation College of Resources and EnvironmentFujian Agriculture and Forestry UniversityFuzhou 350002China State Key Laboratory of Atmospheric Boundary Layer Physics and Atmospheric Chemistry Institute of Atmospheric PhysicsChinese Academy of SciencesBeijing 100029China Guangxi Zhuang Autonomous Region plant protection station Nanjing 530022China Department of Earth Atmosphericand Planetary SciencesPurdue UniversityWest Lafayette IN47907US Department of Bioproducts and Biosystems Engineering University of Minnesota-Twin CitiesSaint Paul MN 55108USA Department of Geography The University of Hong KongHong Kong 999077China

Soil phosphorus (P) plays a vital role in both ecological and agricultural ecosystems, where total P (TP) in soil serves as a crucial indicator of soil fertility and quality. Most of the studies covered in the literature employ a single or narrow range of soil databases, which largely overlooks the impact of utilizing multiple mapping scales in estimating soil TP, especially in hilly topographies. In this study, Fujian Province, a subtropical hilly region along China’s southeast coast covered by a complex topographic environment, was taken as a case study. The influence of the mapping scale on soil TP storage (TPS)estimation was analyzed using six digital soil databases that were derived from 3 082 unique soil profiles at different mapping scales, i.e., 1:50 000 (S5),1:200 000 (S20), 1:500 000 (S50), 1:1 000 000 (S100), 1:4 000 000 (S400), and 1:10 000 000 (S1000). The regional TPS in the surface soil (0–20 cm) based on the S5, S20, S50, S100, S400, and S1000 soil maps was 20.72, 22.17, 23.06, 23.05, 22.04, and 23.48 Tg, respectively, and the corresponding TPS at0–100 cm soil depth was 80.98, 80.71, 85.00, 84.03, 82.96, and 86.72 Tg, respectively. By comparing soil TPS in the S20 to S1000 maps to that in the S5map, the relative deviations were 6.37%–13.32%for 0–20 cm and 0.33%–7.09%for 0–100 cm. Moreover, since the S20 map had the lowest relative deviation among different mapping scales as compared to S5, it could provide additional soil information and a richer soil environment than other smaller mapping scales. Our results also revealed that many uncertainties in soil TPS estimation originated from the lack of detailed soil information, i.e., representation and spatial variations among different soil types. From the time and labor perspectives, our work provides useful guidelines to identify the appropriate mapping scale for estimating regional soil TPS in areas like Fujian Province in subtropical China or other places with similar complex topographies. Moreover, it is of tremendous importance to accurately est

关键词： agricultural management appropriate mapping scale digitized conventional soil map estimation uncertainty subtropical hilly region

New geographic distribution and molecular diversity of Citrus chlorotic dwarf-associated virus in China

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Journal of Integrative agriculture 2022年第1期21卷 293-298页

作者： YANG Zhen ZHANG Lan ZHAO Jin-fa ZHANG Xing-kai WANG Ying LI Tai-sheng ZHANG Wei ZHOU Yan National Citrus Engineering and Technology Research Center Citrus Research InstituteSouthwest UniversityChongqing 400712P.R.China Horticulture Research Institute Guangxi Academy of Agricultural SciencesNanning 530000P.R.China plant protection station of sichuan agriculture and rural bureau Chengdu 610041P.R.China

In 2009, an emerging citrus viral disease caused by Citrus chlorotic dwarf-associated virus(CCDaV) was discovered in Yunnan Province of China. However, the occurrence and spread of CCDaV in other citrus-growing provinces in China is unknown to date. To better understand the distribution and molecular diversity of CCDaV in China, a total of 1 772 citrus samples were collected from 11 major citrus-growing provinces and were tested for CCDaV by PCR. Among these, 134 citrus samples from Guangxi, Yunnan and Guangdong were tested positive for CCDaV, demonstrating that the occurrence and spread of CCDaV are increasing in China. The complete genome sequences of 17 CCDaV isolates from different provinces and hosts were sequenced. Comparisons of the whole-genome sequences of the 17 CCDaV isolates as well as the 15 isolates available in GenBank revealed that the sequence identity was about 99–100%, showing that the CCDaV isolates were highly conserved. Phylogenetic studies showed that the 32 CCDaV isolates belonged to four different groups based on geographical origins and host species, and that CCDaV isolates from China and Turkey were clustered into different groups. The results provide important information for clarifying the distribution and genetic diversity of CCDaV in China.

关键词： Citrus chlorotic dwarf-associated virus detection sequence alignments phylogenetic tree analysis

Possible source and migration pathway for early-summer immigrants of the oriental armyworm, Mythimna separata, arriving in northern Japan

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Journal of Integrative agriculture 2023年第11期22卷 3474-3488页

作者： Akira OTUKA Tokumitsu NIIYAMA JIANG Xing-fu Institute for plant protection National Agriculture and Food Research OrganizationKoshi 861-1102Japan Akita plant protection station Akita 010-1232Japan Institute of plant protection Chinese Academy of Agricultural SciencesBeijing 100193P.R.China

The first generation of the oriental armyworm, Mythimna separata(Walker), arrives every year in northern Japan in mainly late May to early June. Analyses of weather maps suggested that this moth’s immigration source could be eastern China, but the accuracy of those analyses was very limited due to the lack of a current standard trajectory analysis. The management of migratory insect pests such as M. separata benefits from the identification of the migration source(s)and pathway(s) of the pests. The present study provides a trajectory analysis for M. separata. Backward trajectories from trap sites in northern Japan were calculated with the HYSPLIT System developed by the U.S. National Oceanic and Atmospheric Administration, taking the flight speed of M. separata and the limitation of low ambient temperature at flight height into account. The ending times of the moth’s short and long trajectories were set at dusk on the day before and two days before the possible arrival date, respectively. The results suggested two types of possible migration pathway: a multi-step pathway from Northeast China, the Korean Peninsula, and eastern Russia, which are destination areas of the first-generation’s migration, and a direct pathway from seasonal main emigration areas in eastern China such as Jiangsu and Shandong provinces. These findings contribute to our understanding of the migration ecology of M. separata and can be used for the development of methods to predict the migration of this insect.

关键词： migration backward trajectory oriental armyworm first generation East Asia