QTL mapping of adult plant resistance to stripe rust and leaf rust in a Fuyu 3/Zhengzhou 5389 wheat population

Stripe or yellow rust(YR)and leaf rust(LR)cause large losses in wheat production worldwide.Resistant cultivars curtail the levels of losses.The present study aimed to identify quantitative trait loci(QTL)for YR and LR resistance in 147 F2:6 recombinant inbred lines(RIL)derived from the cross Fuyu 3/Zhengzhou 5389.The RIL population and parents were genotyped with the Wheat55 K single nucleotide polymorphism(SNP)array and simple sequence repeat(SSR)markers.All materials were also phenotyped for YR severity at Mianyang in Sichuan province and Baoding in Hebei province in the 2015/2016,2016/2017,and 2017/2018 cropping seasons,and LR severity at Zhoukou in Henan province and at Baoding in 2017/2018.Eleven QTL for YR resistance and five for LR resistance were detected using inclusive composite interval mapping(Ici Mapping).Four of these QTL on chromosomes 1 BL,2 BS,3 AL,and 5 AL conferred resistance to both YR and LR.The QTL on 1 BL was Lr46/Yr29,and that on 7 BL might be Lr68.The QTL on chromosome 2 BS was detected at a similar position to previously detected loci.QYr.hebau-3 AL/QLr.hebau-3 AL,QYr.hebau-5 AL/QLr.hebau-5 AL,QYr.hebau-7 DL,QYr.hebau-4 BS,QYr.hebau-6 DL,and QYr.hebau-2 AS are likely to be new.An SSR marker for QYr.hebau-7 DL was developed and validated in a diverse wheat panel from China,suggesting effectiveness in different genetic backgrounds.These QTL with closely linked SNP and SSR markers could be useful for marker-assisted selection in wheat breeding programs targeting durable resistance to both diseases.

Cloning of PsMYB62 and analysis of cadmium resistant in Potentilla sericea

Potentilla sericea is a heavy metal hyperaccumulator landscaping plant.MYB transcription factors play an important role in regulating plant stress response to adversity.However,there are few studies on MYB transcription factors in stress tolerance in Potentilla sericea.In this study,the PsMYB62 gene was successfully cloned from Potentilla sericea.Methods:Bioinformatic analysis and real-time quantitative PCR(qPCR)methods were used to evaluate this gene.The transgenic A.thaliana were obtained by flower dipping and the gene function was identified by determining physiological indicators under cadmium stress.Results:The open reading frame of PsMYB62 is 942 bp,which encodes 313 amino acids(aa)and belongs to the R2R3 MYB transcription factor.The plant overexpression vector PBI121-PsMYB62-GFP was constructed and successfully transferred into A.thaliana.The relative expression level of PsMYB62 was significantly increased by CdCl_(2),NaCl,ABA,and mannitol treatments.The germination rate of transgenic seeds was higher than those of wild type(WT)and empty vector(EV)under different concentrations of cadmium treatment.Upon treatment with 100μmol·L^(−1)of CdCl_(2)·2.5H_(2)O,the activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT)in the transgenic plants were significantly higher than those in the WT and EV.The contents of H_(2)O_(2),O_(2)·−and malondialdehyde(MDA)in transgenic lines were increased,but lower than those in WT and EV.The expression levels of AtGSH,AtPCS,and AtNAS4 that were related to the regulation of cadmium were increased,but the expression levels of transgenic lines were higher than those of WT and EV.Conclusion:The above results showed that PsMYB62 could be induced by cadmium and could improve the cadmium resistance of plants.

Pre-Breeding Genetic Diversity Assessment of Tomato(Solanum lycopersicum L.)Cultivars Based on Molecular,Morphological and Physicochemical Parameters

Appropriate knowledge of the parental cultivars is a pre-requisite for a successful breeding program.This study characterized fruit yield,quality attributes,and molecular variations of ten tomato cultivars during three consecutive generations under greenhouse conditions.Peto 86,Castle Rock,and Red Star cultivars showed the highest fruit yield(kg/plant),total phenolic compounds(TPC),and sap acidity.Principal component analysis categorized the evaluated fruit yield into three groups based on their quality attributes.A robust positive correlation appeared among traits inside each group.A positive correlation was likewise noticed between the first and the second groups.However,a negative correlation was detected between the first,the second and the third group.Molecular profiling,using seven inter-simple sequence repeat(ISSR)primers,produced 60 loci,including 49 polymorphic loci.The molecular analysis also pinpointed the highest genetic similarity(0.92)between P73 and Moneymaker,while the lowest genetic similarity(0.46)was observed between Castle Rock and Moneymaker.The cultivars P73 and Moneymaker showed the lowest genetic distance(2.24),while the highest genetic distance(5.92)was observed between Super Marmand and Peto86,on the one hand,and between Castle Rock and Moneymaker,on the other hand.The chemical analysis of fruit sap indicated the highest levels of TPC,total flavonoids,anthocyanin,ascorbic acid and total soluble solids in Peto 86 and Castle Rock cultivars.Phylogeny analysis of tomato cultivars based on morphological and molecular attributes indicated four distinct clades.Peto 86,Castle Rock,and Red star cultivars can be recommended for the tomato hybridization breeding programs in the future,with other tomato cultivars as potentially high-yielding parents.

Histopathological Effects of the Protein Toxin from Xenorhabdus nematophila on the Midgut of Helicoverpa armigera

Xenorhabdus nematophila HB310, which is highly virulent for many insects, is symbiotic with Steinernema carpocapsae HB310. Toxin II was obtained using methods such as salting out and native-PAGE from the cells of X. nematophila HB310. The histopathology of toxin II on H. armigera larvae was studied by dissecting an olefin slice of the midgut. The symptoms showed that the histopathology of the H. armigera midgut was similar to that of other novel midgut-active toxins such as the δ-endotoxins from Bacillus thuringiensis, as well as Tca from Photorhabdus luminescens W14. The midgut tissues of H. armigera fourth-instar larvae began to transform after the oral intake of the toxin Ⅱ over 6 h. First, the anterior region of the peritrophic membrane （PM） began to degrade followed by the elongation of the columnar cells. The epithelium decomposed gradually, and the midgut tissues were either loose or disordered. The PM disappeared after 12 h but reappeared after 72 h following transient or sublethal exposure to the toxin Ⅱ. Toxin Ⅱ also directly destroyed in vitro PMs of H. armigera.

Isolation and Characterization of NBS-LRR Class Resistance Homologous Gene from Wheat

One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.

Systemic acquired resistance, NPR1, and pathogenesis-related genes in wheat and barley

In Arabidopsis, systemic acquired resistance （SAR） is established beyond the initial infection by a pathogen or is directly induced by treatment with salicylic acid （SA） or its functional analogs, 2,6-dichloroisonicotinic acid （INA） and benzothiadiazole （BTH）. NPR1 protein is considered the master regulator of SAR in both SA signal sensing and transduction. In wheat （Triticum aesfivum） and barley （Hordeum vulgare）, both pathogen infection and BTH treatment can induce broad-spectrum resistance to various diseases, including powdery mildew, leaf rust, Fusarium head blight, etc. However, three different types of SAR-like responses including acquired resistance （AR）, systemic immunity （SI）, and BTH-induced resistance （BIR） seem to be achieved by activating different gene pathways. Recent research on wheat and barley NPR1 homologs in AR and SI has provided the initial clue for understanding the mechanism of SAR in these two plant species. In this review, the specific features ofAR, Si, and BIR in wheat and barley were summarized and compared with that of SAR in model plants of Arabidopsis and rice. Research updates on downstream genes of SAR, including pathogenesis-related （PR） and BTH-induced genes, were highlighted.

Identification and Molecular Tagging of Leaf Rust Resistance Gene (Lr24) in Wheat

This research was aimed to develop AFLP markers co-segregated with gene Lr24 and validate the using for marker assisted selection （MAS）. An F2 population developed from the cross between the resistant line TcLr24 and the susceptible line Thatcher was tested for resistance to the Puccinia triticina races BGQQ and SHRT using for genetic analysis and molecular marker. A total of 224 AFLP primer combinations were used to test the resistant and susceptible parents, as well as the resistant bulk and the susceptible bulk. Four AFLP markers, P-AGA/M-CTT289 bp, P-AGC/M-CAC1ss bp, P-AGC/M- CAC162 bp, and P-ACG/M-CGC239 bp, were co-segregated with Lr24. The AFLP fragment from the primer combination P- ACG/M-CGC was cloned, sequenced and converted into a STS marker named as ASTS212. Thatcher backgrounded NILs and 115 varieties were examined by using this STS marker and the marker SCS13026oz developed by Gupta. 5R615, 5R616, IR13, and 1R17 were identified and validated to contain gene Lr24. The marker is dominant and may be useful in identification the resistance gene Lr24 in wheat and wheat breeding programs.

Identification of AFLP Markers Linked to Lr19 Resistance to Wheat Leaf Rust

AFLP analyses were carried out on Thatcher, 23 near-isogenic lines and F2 generation of TcLr19 × Thatcher, to develop molecular markers for gene Lr19 resistance to wheat leaf rust. Seven markers linked to Lr19 resistance trait were obtained, which were P-AGT/M-GAG289 bp （3.3 cM）, P-ACA/M-GGT102 bp （4.1 cM）, P-ACA/M-GGT106 bp （4.1 cM）, P-AAC/M-CAG123 bp （4.9 cM）, P-AAC/M-GGT203bp （5.0 cM）, P-ACA/M-GGT290bp （5.7 cM）, and P-ATC/M-GAG293bp （9.6 cM）. All of these specific fragments were isolated from the polyacrylamide gels, reamplified, cloned, and sequenced. The research may facilitate genetic mapping, physical mapping, and the eventual cloning of Lr19.

A SSR Marker for Leaf Rust Resistance Gene Lr19 in Wheat

Microsatellite was carded out in Thatcher, six near-isogenic lines and F2 progeny of TcLr19xThatcher to develop molecular markers for leaf rust resistance gene Lr19. Thirteen primer pairs were screened, of which one primer pair Xgwm44 displayed polymorphsim in the population of TcLr 19, Thatcher, and their F2 generations. One marker closed linked to Lr19 resistance trait was obtained, and was named Xgwm44139bp with the genetic distance 0.9 cM. The research shows that Lr19 has more potential in marker-assisted breeding programs in wheat and provides a step stone for mapping genetic map, physical map and the eventual cloning.

Postulation of Leaf Rust Resistance Genes in Seven Chinese Spring Wheat Cultivars

To detect the leaf rust resistance genes in the 7 Chinese spring wheat clultivars Shenmian 99025, Shenmia 99042, Shenmian 85, Shenmian 91, Shenmian 96, Shenmian 1167 and Shenmian 962, Thatcher, Thatcher backgrounded near-isogenic lines and 15 pathotypes of P. triticina were used for gene postulate at the seedling stage, and 9 of the 15 pathotypes were used in the field tests. Molecular markers closely linked to, or co-segregated with resistance genes Lrl, Lr9, LrlO, Lrl9, Lr20, Lr21, Lr24, Lr26, Lr28, Lr29, Lr32, Lr34, Lr35, Lr37, Lr38, and Lr47 were screened to assist detection of the resistance genes. As results, 4 known resistance genes, including Lrl, Lr9, Lr26, and Lr34, and other unknown resistance genes were postulated singly or in combination in the tested cultivars. Shenmian 85, Shenmian 91, Shenmian 96, Shenmian 962, Shenmian 1167, and Shenmian 99042 are potentially useful for wheat production and breeding programs. The result suggested that combining gene postulation, molecular markers and pedigrees is effective and more accuracy method to know the resistance genes in cultivars.

伊朗吉丁虫科昆虫记述(鞘翅目:吉丁虫科)(英文)

本文记述了伊朗不同地区鞘翅目吉丁虫科昆虫(Buprestidae)的一些种类和分布。共采集和鉴定吉丁虫科昆虫18种,隶属4亚科(Buprestinae,Chrysochroinae,Julodinae,Polycestinae)6属(Agrilus,Anthaxia,Psi-loptera,Julodis,Acmaeodera,Acmaeoderella),并报道了部分种类的寄主植物。

Evaluation of Drought Stress-Inducible W<i>si</i>18 Promoter in <i>Brachypodium distachyon</i>

The rice Wsi18 promoter confers drought-inducible gene expression. This property makes it a useful candidate to drive relevant genes for developing drought resistant traits for different monocot crops. In this study, we showed that the Bradi2G47700 gene, the closest homologue to rice Wsi18, was upregulated in Brachypodium distachyon plants exposed to ABA and mannitol. Wsi18: uidA transgenic B. distachyon plants were produced and then subjected to ABA or mannitol treatment. The expression of uidA in three transgenic lines (line 10, 18 and 37) was significantly upregulated in plants exposed to ABA (fold increases of 5.61 ± 0.98, 2.88 ± 0.75 and 9.13 ± 1.96, respectively) compared to the same transgenic plant lines without treatment. The expression of uidA in two transgenic lines (lines 18 and 37) also showed upregulation when treated with mannitol (fold increases of 4.43 ± 1.07 and 8.47 ± 2.90, respectively) compared to the same transgenic plant lines without mannitol treatment. Moreover, GUS histochemical assay showed increased Wsi18 promoter activity in the leaves and stems of transgenic lines upon treatment with ABA or mannitol. This is the first report of the drought inducible rice Wsi18 promoter being active in B. distachyon which is a model plant for molecular biology research of various monocot plants. Taken together, the results indicate that the Wsi18 promoter and its homologue may be explored as a useful tool for drought stress-inducible gene expression in different monocot crops.

A method to refresh dried larval specimens of Erythraeidae

The article reports a method of making dried larval specimens of Erythraeidae,ectoparasitism of insect,refreshed for taxonomic study.The dried larvae are let in cuvettes with 50%ethanol.The cuvette plat is then put into a beaker with water in appropriate amount,fulling to 1/3-2/3 high of the cuvette plat.The beaker is heated until the water boiling,and then stop to heat.After that,the treated larvae are immersed into a container with ethanol-glycerol solution(50%ethanol:glycerol=20:1),and then the container is warmed in a Water-Bath at 60℃for 2h to‘refresh’the larvae.The refreshed larvae are as mollescent as fresh ones and easy to be prepared for morphologic observation and long-term preservation.

A new species in the genus Diaonidia Takahashi(Hemiptera:Coccomorpha:Diaspididae)from China

A new armoured scale,Diaonidia litsea sp.nov.,is described and illustrated.It was found on Litsea monopetala(Roxb.)Pers.in Hainan,China.This new species can be easily distinguished by:the anterior spiracles each with 2–3 discoidal glands;pygidium with 2 pairs of well-developed lobes,and the area around mouthpart strongly sclerotized.An identification key to the adult females in the genus Diaonidia is also provided.

Cytospora pyri promotes Erwinia amylovora virulence by providing metabolites and hyphae

Bacterial-fungal interactions are widespread in nature.We observed that pear orchards affected by Cytospora pyri(formerly Valsa pyri)were often accompanied with Erwinia amylovora.However,the relationship of the two pathogens was unclear.The objective of this study was to determine whether the synergistic effect exists between E.amylovora and C.pyri.We first analyzed the coexistence frequencies of E.amylovora and C.pyri in pear trees.Virulence of the two pathogens,growth,physical interactions,amylovoran production,and expression of genes for amylovoran biosynthesis were conducted.Our results showed that E.amylovora and C.pyri could coexist on the same lesion and caused much more severe disease.We also found that E.amylovora could physically attach to C.pyri and the expression of amylovoran biosynthesis genes were up-regulated with fungal metabolite treatment.These results indicate that E.amylovora and C.pyri can cooperatively interact,which provides C.pyri with an opportunity to promote bacterial dispersal and production of virulence factor in E.amylovora.

A nano-delivery system expands the insecticidal target of thiamethoxam to include a devastating pest,the fall armyworm

Nano-delivery systems have been applied to deliver various synthetic/botanical pesticides to increase the efficiency of pesticide use and reduce the volumes of pesticides applied.Previous studies have supported the hypothesis that the nanocarriers can help expand the insecticidal target of pesticides to include non-target pests.However,the potential mechanism underlying this interesting phenomenon remains unclear.Herein,a widely applied star polycation(SPc)nanocarrier was synthesized to construct a thiamethoxam(TMX)nano-delivery system.The SPc-based delivery system could promote the translocation of exogenous substances across the membrane of Sf9 cells,increase the cytotoxicity of TMX against Sf9 cells by nearly 20%,and expand the insecticidal target of TMX to include Spodoptera frugiperda(the fall armyworm),with a 27.5%mortality increase at a concentration of 0.25 mg/mL.Moreover,the RNA-seq analysis demonstrated that the SPc could upregulate various transport-related genes,such as Rab,SORT1,CYTH,and PIKfive,for the enhanced cellular uptake of TMX.Furthermore,enhanced cell death in larvae treated with the TMX-SPc complex was observed through changes in the expression levels of death-related genes,such as Casp7,BIRC5,MSK1,and PGAM5.The SPc-based nano-delivery system improved the cellular uptake of TMX and expanded its insecticidal target by adjusting the expression levels of death-related genes.The current study mainly identified the transport and cell death genes related to nanocarrier-based insecticidal target expansion,which is beneficial for understanding the bioactivity enhancement of the nano-delivery system.

An AIE Star Polymer with Enhanced Co-Delivery of Drug and Gene for Synergistic Pest Control

Pesticides,as the most common means of pest managements,have caused a series of problems such as pest resistance and environmental pollution.Aggregation-induced emission(AIE)polymers exhibit great potential in biological applications because their fluorescent intensities significantly enhance in the aggregated state.In this paper,an AIE star polymer including a tetraphenyl ethylene(TPE)core and poly(2-(dimethylamino)ethyl methacrylate)arms is designed and further developed as a multi-functional nanocarrier for agricultural pest control.The nanocarrier shows high water solubility,good size stability and AIE imaging ability.Meanwhile,the twisted AIE core and positively charged polymer arms make the nanocarrier efficiently co-load dinotefuran(DIN)and dsRNA via hydrogen bonds and electrostatic interactions,respectively.The AIE star polymer displays low toxicity and high fluorescence traceability.As a result,the nanocarrier co-loading with DIN and dsRNA exhibits a highly synergistic insecticidal effect with higher pest mortality compared to the separate delivery of DIN and dsRNA.This study develops the AIE star polymer to improve the efficiency of co-delivery of drug and dsRNA and proposes a new strategy toward efficient and synergistic pest control.

Identification of an avirulence gene,avrxa5,from the rice pathogen Xanthomonas oryzae pv.oryzae

Xanthomonas oryzae pv.oryzae,the causal agent of bacterial blight in rice,interacts with rice plants in a gene-for-gene manner.The specificity of the interaction is dictated by avirulence(avr) genes in the pathogen and resistance(R) genes in the host.To date,no avr genes that correspond to recessive R genes have been isolated.We isolated an avrBs3/pthA family gene,avrxa5,from our previously isolated clone p58,which was originally from strain JXOIII.The avrxa5 gene converted the PXO99A strain from compatible to incompatible in rice cultivars containing the recessive xa5 gene,but not in those containing the dominant Xa5 gene.Sequencing indicated that avrxa5,which is highly similar to members of the avrBs3/pthA family,encodes a protein of 1238 amino acid residues with a conserved carboxy-terminal region containing three nuclear localization signals and a transcription activation domain.It has 19.5 34-amino-acid direct repeats,but the 13th amino acid is missing in the fifth and ninth repetitive units.Domain swapping of the repetitive regions between avrxa5 and avrXa7 changed the avirulence specificity of the genes in xa5 and Xa7 rice lines,respectively.This indicates that avrxa5 is distinct from previously characterized avrBs3/pthA members.The specificity of avrxa5 toward recessive xa5 in rice could help us better understand the molecular mechanisms of plant-pathogen specific interactions.