Antixenosis and Antibiosis Resistance in Rice Cultivars against <Emphasis Type="Italic">Chilo suppressalis</Emphasis> (Walker) (Lepidoptera: Crambidae)

The striped stem borer, Chilo suppressalis (Walker) (Lepidoptera: Crambidae), is an important pest afflicting rice in most rice-growing countries in the world. Deliniating the categories of resistance in rice genotypes under field conditions could be helpful in managment of this pest. Two categories of resistance, antixenosis and antibiosis, were examined in ten popular and diverse rice genotypes of different origin that had been selected for their resistance to the striped stem borer in a previous study. Significant differences were found between genotypes for the number of egg masses, number of eggs, preference index, larval and pupal weight, larval development time, larval survival rate, larval mine length, and leaf trichome density. It was found that the rice genotypes Novator, A7801, and Nemat had the more pronounced antixenosis-type resistance, whereas AB1 and Shirodi had better antiobiosis-type resistance. Interestingly, the rice genotype AN-74 for which Nemat is the parental line showed both types of resistance and could be effectively used in an integrated pest management of the rice striped stem borer.     

Transgenic insect-resistant maintainer line (IR68899B) for improvement of hybrid rice

Heterosis has helped to increase rice yield in F₁ hybrids by 15–20% beyond the level of inbred semidwarf varieties. For stable yield performance rice hybrids must also possess genetic resistance to biotic stresses. One of these, stem borer, reduces the expected yield of hybrid rice. The truncated synthetic cryIA(b) gene from Bacillus thuringiensis is known to be effective in controlling stem borer. The development of transformation techniques has provided the technology for incorporating this bacterial gene into the rice genome, which has not been possible by conventional breeding methods. We have introduced a new approach of using a transgenic maintainer line for developing an insect-resistant hybrid rice. An elite IRRI maintainer line (IR68899B) has been transformed with the cryIA(b) gene driven by the 35S constitutive promoter using the biolistic method. The integration and expression of the cryIA(b) gene could be demonstrated through Southern and Western blot analyses that have been carried out so far up to the T₂ generations. Insect bioassay data showed an enhanced resistance to yellow stem borer in the Bt ⁺ transgenic plants. This is the first report of the development of a transgenic maintainer line for use in hybrid rice improvement. Received: 17 December 1997 / Revision received: 23 June 1998 / Accepted: 25 September 1998     

Effects of silicon sources on its deposition, chlorophyll content, and disease and pest resistance in rice

Rice (Oryza sativa L.) was grown in pots with pyridine N-oxide (PNO), 4-morpholino pyridine N-oxide (MNO), and sodium meta silicate as the sources for silicon. Aliquots of these were added in fortnightly intervals to seedlings through anthesis stage. The plants were monitored for plant growth characteristics, chlorophyll content (SPAD values), photosystem 2 activity (variable to maximum fluorescence ratio of dark adapted leaves), and for blast and yellow stem borer resistance. Deposition of silica in the leaves was monitored by scanning electron microscopy and silicon mapping. PNO or MNO application resulted in significant silicon accumulation in leaf bundle sheath cells. Application of PNO and MNO imparted disease and pest resistance by increasing silicon uptake of rice plants.     

Transgenic <Emphasis Type="Italic">indica</Emphasis> rice cultivar ‘Swarna’ expressing a potato chymotrypsin inhibitor <Emphasis Type="Italic">pin2</Emphasis> gene show enhanced levels of resistance to yellow stem borer

Transgenic rice was developed from ‘Swarna’, the most popular indica rice cultivar (Oryza sativa L.) in South East Asia, with a potato chymotrypsin inhibitor gene (pin2) through Agrobacterium-mediated transformation. Four out of nine primary transgenic plants had a single-copy T-DNA insertion while other five plants had two copies. Mendelian pattern of inheritance of the transgene (pin2) was observed in the T₁ generation progeny plants. Whole plant bioassays conducted at both vegetative and reproductive stages and cut stem assays showed enhanced levels of resistance of transgenic rice against yellow stem borer. The transgenic rice lines with plant derived proteinase inhibitor genes would develop into resistant cultivars to fit into resistance breeding strategies as an important component of integrated pest management in rice.     

Rice (<i>Oryza sativa</i> L.) Breeding among Hassawi Landrace and Egyptian Genotypes for Stem Borer (<i>Chilo agamemnon</i> 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.     

Transgenic rice plants with a synthetic cry1Ab gene from Bacillus thuringiensis were highly resistant to eight lepidopteran rice pest species

To fully explore the resistance potential of transgenic rice produced by Agrobacterium-mediated transformation, an elite line KMD1 was assessed for its resistance to eight lepidopteran rice pest species. KMD1 contained a synthetic cry1Ab gene from Bacillus thuringiensis under the control of a maize ubiquitin promoter. It was derived from a commercial japonica Chinese rice variety Xiushui 11, and bred true for both agronomic traits and a cry1Ab gene when the bioassays were done in 1998 in the R₅ generation. The eight lepidopteran pest species were: four Pyralidae species: Chilo suppressalis (striped stem borer, SSB), Scirpophaga incertulas (yellow stem borer, YSB), Cnaphalocrocis medinalis (leaf folder), Herpitogramma licarisalis; two Noctuidae: Sesamia inferens (pink stem borer, PSB) and Naranga anescens; one Stayridae: Mycalesis gotama; and one Hesperiidae, Parnara guttata. In laboratory bioassays, 100% mortality was observed in all insect species when their newly hatched or third-instar larvae were fed KMD1 leaf tissues, whereas only 9.65% of the neonates and none of the third-instar larvae died when fed the leaf tissues of non-transgenic control. Moreover, the leaf area of control tissues consumed in four days by stem borers was 20 to 40 times higher than that of KMD1 tissues, and the area of control tissues eaten by leaf-feeding species was 120 to 180 times greater than that of the transgenic tissues. Under natural infestation, no KMD1 plant was visibly damaged by the SSB, YSB and leaf folder in field evaluation. On the other hand, 80, 9.3 and 88.7% of control plants were injured by SSB, YSB, and leaf folder, respectively. These data disclosed that the transgenic line was highly resistant to a broad spectrum of lepidopteran insect species and could be useful in insect resistance breeding programs.     

Gene cloning and expression of aminopeptidase N and cadherin from midgut of the rice stem borer,Chilo suppressalis

Abstract The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N (CsAPN) and a cadherin (CsCad) were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here (CsCad) showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region (CR7), which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin Cry1Ab. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.     

Constitutive and tissue-specific differential expression of the cryIA(b) gene in transgenic rice plants conferring resistance to rice insect pest

 The truncated chimeric Bt gene, cryIA(b) of Bacillus thuringiensis, driven by two constitutive promoters, 35S from CaMV and Actin-1 from rice, and two tissue-specific promoters, pith tissue and pepcarboxylase (PEPC) for green tissue from maize, was introduced into several varieties of rice (indica and japonica) by microprojectile bombardment and protoplast systems. A total of 1800 putative transgenic Bt rice plants could be produced. Southern analysis revealed that more than 100 independently transformed plants could be confirmed for integration of the cryIA(b) gene. High levels of CryIA(b) proteins were obtained in the green tissue (leaves and stem) of many plants using the PEPC promoter. There was little difference in Bt protein level in leaves and stems from transgenic plants with the 35 S or Actin-1 promoter. Out of 800 Southern-positive plants that were bioassayed, 81 transgenic plants showed 100% mortality of insect larvae of the yellow stem borer (Scirpophaga incertulas). The transgene, cryIA(b), driven by different promoters showed a wide range of expression (low to high) of Bt proteins stably inherited in a number of rice varieties with enhanced yellow stem borer resistance. This first report of transgenic indica Bt rice plants with the PEPC or pith promoter either alone or in combination should provide a better strategy for providing rice plants with protection against insect pest resistance, minimizing the expression of the CryIA(b) protein in seeds and other tissues. Received: 12 November 1997 / Accepted: 25 November 1997     

转Cry1Ab和Cry1Ac融合基因型抗虫水稻对田间二化螟和大螟种群发生动态的影响

以Bt水稻华恢1号(Cry1Ac和CryAb融合基因;简称HH1)及其对照亲本明恢63(简称MH63)稻田靶标害虫二化螟Chilosuppressalis和次靶标害虫大螟Sesamia inferens为研究对象,研究了转基因抗虫水稻大田螟虫发生规律及其靶标和次靶标害虫致害力差异。结果表明,Bt水稻及其对照亲本上二化螟或大螟的卵块数量差异不显著,同时,对照亲本上二化螟与大螟的落卵量差异不显著,但Bt水稻上二化螟的落卵量显著大于大螟。与对照亲本相比,Bt水稻上二化螟幼虫发生量显著降低,降幅高达84.9%—100%,但大螟发生量差异不显著;此外,对照亲本上二化螟幼虫发生量显著高于大螟,但Bt水稻上两者差异不显著。同时,Bt水稻上二化螟导致的枯心/白穗率和受害丛率都显著低于其在对照亲本上的致害程度,降幅分别为30.8%—98.3%和11.4%—96.6%,而大螟差异不显著。可见,Bt水稻对靶标害虫二化螟具有较高抗性,而对次靶标害虫大螟的抗性不明显。另一方面,Bt水稻和对照亲本上二化螟导致的枯心/白穗率和受害从率都显著高于大螟。可见,二化螟仍是当前非转基因水稻上的主要害虫,而Bt水稻对二化螟幼虫发生的显著抑制作用以及对大螟幼虫发生的不显著影响,使得其大面积商业化种植下靶标害虫二化螟和次靶标害虫大螟间的竞争替代成为可能。     

Inhibition of midgut protease of yellow stem borer (<Emphasis Type="Italic">Scirpophaga incertulas</Emphasis>) by cysteine protease-like inhibitor from mature jackfruit (<Emphasis Type="Italic">Artocarpus heterophyllus</Emphasis>) seed

A novel protease inhibitor was isolated and purified from the mature seeds of jackfruit (Artocarpus heterophyllus) by precipitation with ammonium sulphate, followed by DEAE-cellulose and gel filtration (Sephadex G-100) chromatography. The isolated protease inhibitor strongly inhibited papain and midgut proteases of yellow stem borer (Scipophaga incertulas) larvae, as seen by in vitro assay. The purified protease inhibitor was active over a wide range of pH with the maximum activity between pH 4 and 10. This protein was also stable up to 80°C, but the retained activity was lost at 100°C, when heated for 30 min. The molecular mass of the purified cysteine-like protease inhibitor is to be 14.50 kDa as determined by SDS-PAGE. Significant reduction in larval weight and mortality was observed, when fresh rice culms with protease inhibitor was feeded to the yellow stem borer larvae. These results may provide important information to control the yellow stem borer in rice with respect to naturally occurring insecticidal proteins. The observed differences would potentially translate into reductions in population growth of yellow stem borer, indicating a potential value of using jackfruit protease inhibitor for protecting rice plants against damage by the yellow stem borer.     

我国稻螟灾害的现状及损失估计

我国稻螟为害的确切记载始于 1 0 2 2年。 2 0世纪 5 0年代中期以前 ,以二化螟Chilosuppressalis(Walker)为害为主。此后十几年中 ,三化螟Tryporyzaincertulas(Walker)普遍大发生。 70年代 ,我国螟害轻微。 1 993年 ,二化螟急剧回升 ,三化螟也随之回升。 1 996年 2种稻螟大发生 ,此后连年暴发成灾。目前我国螟灾的主要特点是虫口密度高、受害范围广、受害程度重、持续时间长、经济损失大。首要螟灾区是沿江稻区 ,二化螟和三化螟分别约占 2 3和 1 3。全国年发面积约 1 5 0 0万hm2 ,防治约 3 80 0万hm2 次。该文首次估计年防治代价约 45 7～ 60亿元 ,残虫造成作物损失近 65亿元 ,总经济损失约 1 1 5亿元。由此可见 ,当前稻螟已取代稻飞虱、棉铃虫成为影响我国国计民生的头号害虫。     

New insights into an RNAi approach for plant defence against piercing‐sucking and stem‐borer insect pests

Insect double‐stranded (ds)RNA expression in transgenic crops can increase plant resistance to biotic stress; however, creating transgenic crops to defend against every insect pest is impractical. Arabidopsis Mob1A is required for organ growth and reproduction. When Arabidopsis roots were soaked in dsMob1A, the root lengths and numbers were significantly suppressed and plants could not bolt or flower. Twenty‐four hours after rice roots were immersed in fluorescent‐labelled dsEYFP (enhanced yellow fluorescent protein), fluorescence was observed in the rice sheath and stem and in planthoppers feeding on the rice. The expression levels of Ago and Dicer in rice and planthoppers were induced by dsEYFP. When rice roots were soaked in dsActin, their growth was also significantly suppressed. When planthoppers or Asian corn borers fed on rice or maize that had been irrigated with a solution containing the dsRNA of an insect target gene, the insect's mortality rate increased significantly. Our results demonstrate that dsRNAs can be absorbed by crop roots, trigger plant and insect RNAi and enhance piercing‐sucking and stem‐borer insect mortality rates. We also confirmed that dsRNA was stable under outdoor conditions. These results indicate that the root dsRNA soaking can be used as a bioinsecticide strategy during crop irrigation.     

Composition of stem borer communities in selected vegetation mosaics in Kenya

Busseola fusca (Fuller), Sesamia calamistis Hampson, Chilo partellus (Swinhoe) and Chilo orichalcociliellus (Strand) are important stem borer pests of maize and sorghum in East Africa. Persistence of these pests in crop fields is blamed on the influx of diaspore populations from the neighbouring natural habitats. In addition to pest species, natural habitats support numerous non-economic stem borer species, some not known to science. However, due to growing human populations and accompanying global change, some of the natural habitats are undergoing rapid changes, a process that may result in the evolution of “new” pest species. In this study, we investigated stem borer species diversity in four different vegetation mosaics in Kenya, with an aim of establishing the differences in species composition and distributions in both wild and cultivated habitats. We identified 33 stem borer species belonging to 14 different genera in the four families; Noctuidae, Crambidae, Pyralidae and Tortricidae from 37 plant species. In addition to the above stem borer pest species, we found three more species, Busseola segeta Bowden, Pirateolea piscator Fletcher and Eldana saccharina Walker, in the cultivated fields. Together, stem borer pests varied in distribution among vegetation mosaics, suggesting differences in ecological requirement. Despite the variations in distribution patterns, stem borer pests co-existed with non-economic species in the natural habitats, communities that are facing threats due to ongoing habitat changes. This paper discusses the likely impacts of habitat changes on both pest and non-economic species.     

Biochemical and enzymatic changes in rice plants as a mechanism of defense

A laboratory study was undertaken to ascertain the impact and the extent of feeding by different pests on biochemical constituents and various enzyme levels in rice plants. The difference in these parameters due to the pest damage by three different modes of feeding was also studied and compared. The borer pest—yellow stem borer (YSB), Scirpophaga incertulas (W); surface feeder—-leaf roller (LR), Cnaphalocrosis medinalis (G) and a sucking pest—brown plant hopper (BPH), Nilaparvata lugens (S) fed rice plants were analyzed for the quantitative and qualitative changes in biochemical profile and enzymatic changes that occur as plant’s defensive responses were analyzed spectrophotometrically. The phenolic acids were analyzed using HPLC and quantitated with the standard samples. The quantity of biochemicals such as proteins, phenols and carbohydrates has been enhanced along with the enzyme activities of peroxidase (POD), catalase (CAT), chitinase (CHI). A decrease in superoxide dismutase (SOD), phenyl alanine ammonia lyase (PAL), β-1, 3-glucanase (GLU) enzyme activities were evident in pest infested plants. Phenolic acids like vanillic acid, syringic acid, cinnamic acid, and p-coumaric acids were mostly found in the infested plants. We demonstrate that the elevated levels of biochemicals, phenolic acids, and enzymes may play a major role in plant defense.     

Gene cloning and expression of aminopeptidase N and cadherin from midgut of the rice stem borer,Chilo suppressalis

The rice stem borer,Chilo suppressalis Walker is one of the most importantinsect pests on rice in Asia,north Africa and southern Europe.Transgenic Bt rice has beendeveloped in the laboratory with good resistance to this pest and other Lepidopteran insects,which will provide a possible alternative tool for this pest control.The full-length cDNAsencoding an aminopeptidase N (CsAPN) and a cadherin (CsCad) were cloned from C.suppressalis.CsAPN showed common features of,and high identities to,other insect AP...     

Development of Transgenic Sugarcane Resistant to Sugarcane Borer

Sugarcane borer (Diatraea saccharalis) is a major pest of sugarcane (Saccharum spp. hybrids), across the Americas. The insect is partially controlled by biological and chemical means, but still causes significant economic losses to sugarcane growers and processors. Proteins derived from Bacillus thuringiensis, Bt proteins, have been used to control sugarcane borer in maize (Zea mays) for the past decade. In sugarcane, the expression of individual Bt proteins has been reported several times. However practical use of Bt proteins requires their use as part of an Integrated Pest Management (IPM) system that includes the delivery of high doses of protein and the use of a refuge to slow the evolution of insect resistance to the protein. Here we demonstrate the feasibility of using Bt proteins to protect sugarcane from sugarcane borer in a commercial setting. We have expressed two Bt proteins with differing modes of action (Cry1Ab and Cry2Ab) in commercial sugarcane varieties, demonstrated efficacy against sugarcane borer in the field and describe a strategy for trait deployment in this tropical crop with complex genetics that limits trait introgression by backcrossing.     

Specificity of systemically released rice stem volatiles on egg parasitoid,Trichogramma japonicum Ashmead behaviour

The yellow stem borer, Scirpophaga incertulas Walker (YSB), infested rice plants emit chemicals through the surface of their infested stems. These induce attractant activity and cause arrestment responses and ovipositional stimulation in its egg parasitoid, Trichogramma japonicum Ashmead. Laboratory experiments on short‐range host searching and oviposition were performed to assess the how these crude stem extracts and their fractions influence the biological control efficiency of Trichogramma. The activity of these chemicals as long‐range attractants was confirmed through wind tunnel bioassays. Stem borer‐infested plant extracts had enhanced the parasitization rate of T. japonicum, whereas host eggs treated with the extract from undamaged stems or solvent‐treated control failed to evoke changes in the parasitoid’s behaviour. A preliminary GC‐MASS analysis indicated the presence of several hydrocarbon compounds. The analysis also revealed qualitative and quantitative differences between the chemical profiles of the infested and non‐infested plants. We hypothesized that herbivore‐induced plant chemicals are released through the stem surfaces and attract T. japonicum, even over long distances. These cues elicit parasitoid arrestment on pest‐damaged plants and subsequently lead to the successful parasitization of the stem borer.     

月相对两种稻螟灯下诱蛾量的影响

经圆形统计分析安徽省庐江县 1 962～ 2 0 0 1年灯光诱蛾资料 ,发现三化螟和二化螟的诱蛾量受到月相的显著影响。诱蛾量的平均角或集中趋势 ,三化螟对应于农历月 2 7日 1 0时 1 3分 (P <0 0 0 1 ) ,二化螟对应于 2 4日 1 6时 2 2分 (P <0 0 0 1 )。两种稻螟诱蛾量的集中时间差异不显著 (P >0 0 5 )。二者合并 ,得到平均集中时间为 2 5日 1 4时 40分。     

两种稻螟灯下蛾峰日与月相的关系

经常规及圆形统计分析我国 2 0世纪 5 0年代灯光诱蛾资料 ,发现三化螟和二化螟的蛾峰日受到月相的显著影响。三化螟蛾峰数的平均角 (集中趋势 )对应于农历 2 6日 0时 5 3分 (P<0 0 0 1) ,二化螟对应于 2 4日 2 3时 5 3分 (P <0 0 0 1) ,二者差异不显著 (P >0 2 5 ) ,合并对应于 2 5日 11时 36分。     

Performance and preference of sugarcane borer,Diatraea saccharalis,on rice cultivars

The sugarcane borer, Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae), is a major agronomic pest in the southeastern USA. This study was conducted to investigate the performance of D. saccharalis on eight rice cultivars widely grown in Louisiana and to assess the relationship between oviposition preference and offspring performance. In a previous study, oviposition preference of sugarcane borers was found to vary on the same eight rice cultivars. In this study, variation in larval performance on the eight cultivars was observed. The overall performance of larvae in terms of relative growth rate and boring success of larvae was highest on Priscilla and Cocodrie. Larval performance was lower on the hybrid XL723, the Clearfield (herbicide‐resistant) variety CL161, and the medium grain Bengal. Also, larvae took more time to enter into the stem of the hybrid XL723 compared with Cocodrie. Significant positive correlations were observed among relative growth rate and boring success and between larval performance and oviposition preference. Significant positive correlations among measures of larval performance and oviposition preference suggest the operation of a common resistance mechanism. Results in laboratory and greenhouse extended into the field, where Cocodrie and Cheniere were the most damaged cultivars, whereas CL161 and the medium grain Jupiter were least injured in terms of average number of stem borer entry/exit holes per plant. Results from this study could contribute to the use of resistant cultivars in an integrated management program for stem borers and the resistant cultivars identified in this study could be used as sources of resistance in breeding programs for resistance against stem borers.