Transgenic japonica rice expressing the cry1C gene is resistant to striped stem borers in Northeast China

Rice production and quality are seriously affected by the lepidopteran pest,striped stem borer(SSB),in Northeast China.In this study,a synthetic cry1 C gene encoding Bacillus thuringiensis(Bt)δ-endotoxin,which is toxic to lepidopteran pest,was transformed into a japonica rice variety(Jigeng 88)in Northeast China by Agrobacterium-mediated transformation.Through molecular detection and the Basta resistance germination assay,a total of 16 single-copy homozygous transgenic lines were obtained from 126 independent transformants expressing cry1 C.Finally,four cry1 C-transgenic lines(JL16,JL23,JL41,and JL42)were selected by evaluation of the Cry1 C protein level,insect-resistance and agronomic traits.The cry1 C-transgenic lines had higher resistance to SSB and higher yield compared with non-transgenic(NT)control plants.T-DNA flanking sequence analysis of the transgenic line JL42 showed that the cry1 C gene was inserted into the intergenic region of chromosome 11,indicating that its insertion may not interfere with the genes near insertion site.In summary,this study developed four cry1 C-transgenic japonica rice lines with high insect resistance and high yield.They can be used as insect-resistant germplasm materials to overcome the problem of rice yield reduction caused by SSB and reduce the use of pesticides in Northeast China.

Field Efficacy Test of 22% Fipronil FS against Sugarcane Stem Borers and Thrips

[ Objective] The paper was to explore the field efficacy of 22% fipronil FS against sugarcane stem borer and thrips （Baliothrips serratus Kobus ）. [ Method] The field trial was conducted by foliar spraying of 22% fipronil FS and mixed broadcasting with fertilizer in 2014. [ Result ] Foliar spraying of 22% fipronil FS at the dose of 50 mIV667 m~ had the best control effect against sugarcane stem borer, and the control effect at 90 d post administration was 70.70%, significantly higher than that of control agent （61.86%）. Medium and high dose of 22% fipronil FS had better control effects against sugarcane thrips. [ Conclusion ] Considering control effects against sugarcane stem borers and thrips as well as sugarcane yield, it is recommended that 22% fipronil FS is sprayed at medium concentration （50 mL/667 m2 ）.

Differential effects of insecticides on mitochondrial membrane fluidity and ATPase activity between the wolf spider and the rice stem borer

Differential effects of methamidophos and three pyrethroids on ATPase activity and membrane fluidity of mitochondria were investigated between the wolf spider（Pirata subpiraticus（Boes.et Str.））and the rice stem borer（Chilo suppressalis（Walker））.Based on a comparison of LD_（50） values,the toxicities of the tested insecticides were higher to the wolf spider than to the rice stem borer.Cyhalothrin at 1×10^（–4） mmol L^（–1） caused inhibition of the mitochondrial Na~＋-K~＋-ATPase and Ca^（2＋）-Mg^（2＋）-ATPase activities,and it’s inhibitions on Na＋-K＋-ATPase and Ca^（2＋）-Mg^（2＋）-ATPase activities were significantly higher in the wolf spider（44 and 28%）than in the rice stem borer（19 and 11%）.Methamidophos at 1×10^（–4） mmol L^（–1） decreased Ca^（2＋）-Mg^（2＋）-ATPase activity by 16 and 27%in the wolf spider and the rice stem borer,respectively,but no significant effect on the specific activity of Na＋-K＋-ATPase was observed.The DPH（1,6-diphenyl-1,3,5-hexatriene）fluorescence polarization values of mitochondrial membranes were not significantly affected by methamidophos in either species.However,cyhalothrin and alpha-cypermethrin induced the values of DPH polarization of mitochondrial membrane increasing with the concentration of cyhalothrin and alpha-cypermethrin from 20 to 100μmol L^（–1） in the rice stem borer and the wolf spider.Effect of ethofenprox on fluidity of the wolf spider and the rice stem borer was contrary.These results suggest that both inhibition of membrane ATPase and changes of membrane fluidity could be appended to the action mechanisms of pyrethroid insecticides.

The insecticide resistance in stripped stem borer,Chilo suppressalis(Walker)

The stripped stem borer (SSB),Chilo suppressalis (Walker) is one ofthe major insect pests of rice in Chi-na. Chemical control has been acommon practice in SSB managementsince 1950s. Insecticides used in-cluded BHC before 1983; organophos-

Rice(Oryza sativa L.)Breeding among Hassawi Landrace and Egyptian Genotypes for Stem Borer(Chilo agamemnon Bles.)Resistance and Related Quantitative Traits

Rice stem borer(Chilo agamemnon Bles.)is a primary insect pest of rice and is a major limiting factor to rice production.Breeding for insect-resistant crop varieties has been an economic way of integrated pest management(IPM)as it offers a viable and ecologically acceptable approach.This study was aimed to evaluate rice genotypes for their resistance against rice stem borer.Seven parental genotypes with twenty one F1 crosses were evaluated for genotypic variation in field experiments.Analysis of variance revealed significant differences for the studied traits in almost all crosses and parents.In addition,the mean squares of parents versus their crosses were signifi-cant for stem borer resistance and other associated traits.Moreover,both general combining ability(GCA)and specific combining ability(SCA)variances were highly significant for all characters studied in the F1 generation.Based on GCA,4 genotypes(Sakha101,Gz6903-3-4-2-1,Gz9577-4-1-1 and Hassawi)exhibited highly significant negative values for stem borer resistance(–0.53,–1.06,–0.18 and–0.49,respectively)indicating they are the best combiners for stem borer resistance.Based on SCA analysis,nine cross combinations showed highly significant negative effects for stem borer resistance.Similarly,the cross Giza178
Hassawi was the best combination with significantly highest value for early maturity.In addition,seven crosses showed highly significant negative SCA for plant height trait.On the other hand,for panicle length,number of primary branches/panicle,panicle weight and 1000-grain weight,seven,four,eight and six crosses showed highly significant positive SCA,respectively.The result further revealed that the non-additive dominance genetic variance was higher than the additive variance for all evaluated traits indicating that non-additive genetic variances have a role in their inheritance.The broad-sense heritability estimates were high for all the studied traits.The stem borer resistance was significantly correlated with panicle weight and 1000-grain weight,which also showed a highly significant correlation with grain yield/plant.Thus these traits can be effectively employed in a breeding program to confer resistance against stem borer infestation in rice.It was further supported by biplot analysis,which clustered these potentially important traits into two quadrants showing their importance in any future breeding program to control stem borer infestation.This study has contributed valuable information for evaluation of genetic diversity in the local rice germplasm and its utilization in futuristic rice genetic improvement programs.

Identification of Bt-transgenic Rice Plants for Resistance to Stripe Stem Borer (Chilo suppressalis) and Genetic Analysis of the Transgenes

Resistance to herbicide Basta was used to identify Bt-transgenic rice plants and the progenies of crosses between the Bt-transgenic rice and the rice varieties cultivated in the Huang Huai area of China. The results demonstrated that the Basta-positive rice plants were highly resistant to stripe stem borer (Chilo sup-pressalis) both in the laboratory and field tests. Both cryIA (b) and bar genes were expressed and co-inherited in both self ing and crossing progenies. Mendelian segregation of the marker gene bar was observed in F2 and BC1 progenies. The results implicates that it is possible to transfer cryIA (b) gene into other cultivated varieties through crosses and back crosses.

Study of Information Management System for the Population of Yellow Stem Borer [Tryporyza incertulas(walker)] Based on Event Ontology

Based on the representation method of conventional knowledge, a new knowledge representation method for population of yellow stem borer [Tryporyza incertulas（walker）]based on event ontology is proposed, and the new information management system for the population of yellow stem borer is constructed based on the method of knowledge representation. Compared with traditional knowledge representation, representation method of event ontology knowledge adopts higher granularity events as knowledge representation units, which is more accorded with objective reality, having better scalability and integration.

Outbreak, Distribution and Damage Characteristics of Cocoa Stem Borer, Eulophonotus myrmeleon Felder 1874 (Lepidoptera: Cossidae) in Major Cocoa Producing States in Nigeria

This study aims to ascertain the extent of the cocoa stem borer Eulophonotus myrmeleon distribution, outbreak, and damage to the cocoa plantations in six cocoa growing states in Nigeria. Three local government areas were selected from each of the six major cocoa states and three farms from each local government area. In each farm, 100 cocoa trees were randomly selected and inspected for symptoms and damages of E. myrmeleon. The result revealed that E. myrmeleon infestation has increased significantly (p = 0.01, df = 1, F = 41.77) in 2019 by 10% from the 2014 inspection. It also showed that the infestation level was significantly different from state to state. Symptoms of infestation include presence of entry/exit holes, woody exudates around the trees, yellowing and browning of the leaves. Damages caused by E. myrmeleon infestation are, loses of pod on the affected branch and tree and death of tree. Field observations showed clearly that stem borer is now becoming a serious problem and limiting considerably the production of cocoa in the region. It is time to give more attention to the biology and management of the pest. This is the first report detailing the symptoms and damage done by cocoa stem borers in Nigeria.

Damage and economical threshold of yellow stem borer (Tryporyza incertulas) on rice

Systematic measurements to damage and yieldloss due to yellow stem borer (YSB) weremade in the single crop and double crop areasof Hangzhou, Jiaxing, and Huzhou cities,northern Zhejiang Province in 1987-1994. YSBeggmasses were released on rice plants at 0,1, 3, 6, 9, 12 eggmasses per plot (equal to0, 495, 1 500, 3 000, 4 500, and 6 000eggmasses per hectare) from tillering to head-ing. The plot size was 20 m~2. There were 3replications randomly lined. The results in 6locations during 4-6 years of 1987-1994showed that the grain losses caused by eacheggmass at tillering stage was 59.11±2.99 g,elongating 63.18±1.51 g, and heading 74.58

Mechanism of rice variety resistance to the yellow stem borer,Scirpophaga incertulas (Walker)

Yellow stem borer(YSB),Scir-pophaga incertulas(Walker)has be-come the main rice insect pest onceagain along the Changjiang Riversince 1990s.Unfortunately,no ricevariety with resistance to YSB hasbeen reported so far.Japonica Zhen-dao 2 with moderate resistance toYSB was found.In 10 rice vari-eties with different resistance levels,YSB individuals showed distinct de-velopmental rates,and the asyn-chronous development even occurredin the same rice variety.In this

Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene

A synthetic cry2A^＊ gene enco ding Bacillus thuringiensis（Bt） δ-endotoxi n that resi st ance to lepidopteran pest was transformed into japonica rice variety Jijing 88, which is the most widely cultivated variety in Jilin Province, Northeast China, by Agrobacterium-mediated transformation. A total of 106 independent transformants overexpressing cry2A^＊ gene driven by ubiquitin（Ubi） promoter was produced. Three single-copy homozygous transgenic lines were finally selected based on the results of PCR analysis, se gregation ratio of Bast a resistance, and Southern hybridiza tion analyse s. RT-PCR and enzyme linke dimmune sorbent assay（ELISA） revealed that cry2A^＊ transcripts and protein were highly expressed in these lines. The high level of Cry2A^＊ protein expression resulted in high resistance to rice striped stem borer as evidence d by insect feeding bioassays. Our results demonst rate that cry2A^＊ transgenic japonica rice confers resistance to the rice striped stem borer in the laboratory conditions.

Rice gene expression profiles responding to larval feeding of the striped stem borer at the 1st to 2nd instar stage

The objective of this study was to identify rice genes that are in response to the striped stem borer （SSB） （Chilo suppressalis Walker） feeding at the first to second larval stage. Using combined suppression subtractive hybridization （SSH） and dot blot approaches, we analyzed the induced defense genes that took place during the first 72 h of infesting intact rice （Oryza sativa L.） plants in sheath tissues with SSB larvae. By sequencing the whole SSH library, 39 expressed sequence tags involved in disease stress, insect stress or other stress responses were identified to be up-regulated by SSB larvae feeding. Among these genes, rice allene oxide cyclase （AOC）, terpene synthase （TPS） and four proteinase inhibitor （PI） genes were up-regulated by SSB larvae feeding. Real-time quantitative reverse transcription polymerase chain reaction analysis showed that four rice PI genes were already up-regulated at 6 h, and reached peaks between 6 h to 12 h. In addition, the transcription ofgene involving in jasmonate signaling pathway such as allene oxide cyclase （AOC） concerning rice early defense response to SSB feeding was activated after rice feeding by SSB for 2 h. Although the expression office terpene synthase （TPS） gene, involved in the biosynthesis ofmonoterpenes or diterpenes, was already up-regulated at 7 h, a significant increase in the expression was delayed until 12 h and reached its peak at 24 h. The present study identified six SSB-response genes and their expression patterns, which provides evidence and information to understand insect stress-response in plants.

Cloning of a Potato Proteinase Inhibitor Gene PINII-2x from Diploid Potato(Solanum phurejia L.) and Transgenic Investigation of Its Potential to Confer Insect Resistance in Rice

Both cDNA and a genomic DNA fragment encoding a new potato proteinase inhibitor Ⅱ were isolated from a diploid potato IVP101 （Solanum phurejia L.） and named PINII-2x. Nucleotlde sequencing confirmed that the DNA fragment of PINll-2xwas 580 bp, including a 115-bp intron and two exons. The deduced PINII-2x proteln contained an Intact signal peptide and two active sites. The PINII-2x gene and its deduced PINII-2x protein had 88% and 93% homology with another tetraploid potato proteinase inhibitor Ⅱ, respectively. Northern blotting analysis Indicated that the mRNA of PINII-2x gene was wound induced in potato leaves. Binary vector pNAR301 and pNAR302 were constructed for rice transformation, in which the PINII-2x cDNA was driven, respectively, by rice actin I promoter （Actl） and maize ubiquitin promoter （Ubll）. Via an Agrobacteriummediated method, these two constructs were transferred into japonica rice cv. Xiushui 63. PCR and Southern blotUng analysis for transgenic rice revealed the integration of the PINII-2x gene. Northern blotting analysis also confirmed transcripts of the PINII.2x gene in transgenic rice plants. Insect bloassays using stripe stem borer （Chilo auppressalis Walker） demonstrated that the average weight and body length of larvae In transgenic plants were only nearly 50% and 61% of those of larvae in control plants, respectively. These results Indicate that the PINII-2x gene should be an effective insect-resistance gene and could be valuable for application in crop breeding for insect resistance.