A binding protein for fungal signal molecules in the cell wall of Pisum sativum

In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules.     

Localization and responsiveness of a cowpea apyrase VsNTPase1 to phytopathogenic microorganisms

Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean, and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli. In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria). A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of apyrases in recognizing and responding to exogenous signals. The nucleotide sequences of VsNTPase1 and VsNTPase2 in this article have been submitted to DDBJ as accession numbers AB196769 and AB196770, respectively.     

Addition of <Emphasis Type="Italic">Pestalotiopsis </Emphasis>spp. to leaf spot pathogens of Japanese persimmon

 In June 1996, a leaf spot disease widely occurred in Japanese persimmon (Diospyros kaki) orchards in Tottori Prefecture, Japan. The main diagnostic symptom was ring spot on the leaves and calyxes of young fruits; in severe cases, lesions developed on more than half of the area of the leaf, resulting in early defoliation. Based on morphological and pathological studies of the isolated fungi, it was shown that Pestalotiopsis longiseta, P. glandicola, P. acaciae, and P. crassiuscula were responsible for the diseases. These fungi, except P. longiseta, were found to be new pathogens of the disease. Received: May 20, 2002 / Accepted: July 25, 2002     

Aster yellows subgroup (Candidatus Phytoplasma sp. AY 16S-group,AY-sg) phytoplasma associated with porcelain vine showing witches' broom symptoms in South Korea

 This is the first report of a phytoplasma in porcelain vine [Ampelopsis brevipedunculata var. heterophylla (Thunb.) Hara.] with severe witches' broom symptoms in Korea. On the basis of the polymerase chain reaction-amplified ribosomal DNA, the phytoplasma infecting porcelain vine was classified as a member of the aster yellows subgroup. Received: October 21, 2002 / Accepted: December 20, 2002     

Role of the α subunit of heterotrimeric G-protein in probenazole-inducing defense signaling in rice

 Certain cellular responses to stresses and stimuli are regulated by a G-protein-mediated signaling pathway. A rice dwarf mutant that is defective in the α subunit of the heterotrimeric G-protein was found to be fully protected from blast fungus by the plant activator probenazole (PBZ) despite the 1-day delay in induction of the PR-10 gene PBZ1 by PBZ. These results suggest that the signaling pathway for protection by PBZ is not via the G-protein, although G-protein is involved in the induction of PBZ1 by PBZ. Received: March 27, 2002 / Accepted: August 20, 2002     

Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Leaf spot of <Emphasis Type="Italic">Aster savatieri</Emphasis> (Makino) Kitam. caused by <Emphasis Type="Italic">Septoria chrysanthemella</Emphasis> Saccardo

 In May 1998 leaf spot caused by Septoria chrysanthemella was found on Aster savatieri in Kanagawa Prefecture, Japan. This is the first report of leaf spot on A. savatieri caused by S. chrysanthemella. Received: September 13, 2002 / Accepted: October 18, 2002 Acknowledgments The authors thank Dr. T. Kobayashi, formerly of Tokyo University of Agriculture, for his advice on identifying the fungus.     

椴树科树种抑虫机制的研究

通过调查研究发现 ,在不同立地条件下 ,光肩星天牛 ( Anolophora glabripennis)在椴树上卵的孵化率不尽相同 ,它与椴树韧皮部厚度、刻槽大小、卵的位置及生长速度没有直接的关系。椴树枝干的内含物直接影响着天牛幼虫的发育和存活     

Involvement of carbon catabolite repression on regulation of endopolygalacturonase gene expression in citrus fruit

 The Acpg1 gene of Alternaria citri encodes an extracellular endopolygalacturonase that is important for virulence in citrus fruits. Expression of Acpg1 is regulated by substrate induction and carbon catabolite repression. A green fluorescent protein (GFP) gene was employed as a reporter gene to define 813 bases upstream of the translation start site comprising the Acpg1 promoter. This upstream sequence contains five putative binding sequences of catabolite repressive element A (CreA), a cis-acting zinc finger repressor involved in carbon catabolite repression. We constructed each CreA-binding site-deleted Acpg1 promoters with GFP reporter gene and transformed them to A. citri. The construct PGPDL4 deleted from −401 to −813 showed both substrate induction and catabolite repression, whereas PGPDL5 additionally deleted from −1 to −84, including one putative CreA-binding site, resulted in a loss of catabolite repression function. Green fluorescence of PGPDL4 was induced by pectin in the peel but was repressed completely in the juice sac area of citrus fruit. However, green fluorescence of PGPDL5 was induced in both the peel and juice sac area, indicating that repression of Acpg1 in the juice sac area is likely accomplished by carbon catabolite repression. Received: October 4, 2002 / Accepted: October 31, 2002 Acknowledgments The authors thank Drs. D. Cullen, A. Van den Wymelenberg, and J. Andrews, University of Wisconsin, for providing pTEFEGFP containing GFP and Dr. T. Tsuge, Nagoya University, for providing transformation vector pSH75. The nucleotide sequence data of Acpg1 promoter region in the DDBJ, EMBL, and GenBank sequence databases is under accession number AB047543. This research was supported in part by grants to K.A. from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.     

泽漆乳浆及乙醇提取物对桃蚜的生物活性研究

使用浸渍法测定了泽漆乳浆及乙醇提取物对桃蚜的室内杀虫作用和拒食作用。结果表明其乳浆物浓度在9.77g/L对桃蚜24h后校正致死率为75.39%,48h后校正致死率为86.08%;其乙醇提取物浓度20g/L24h的校正致死率为60.29%,48h后桃蚜校正致死率为63.06%。通过对泽漆乳浆和乙醇提取物杀虫效果和拒食作用的回归分析,表明不同浓度泽漆乳浆与提取物的触杀作用和拒食作用呈线性关系,且在浓度较大时,乳浆对桃蚜的致死率高于提取物,表明乳浆中对蚜虫致死物质的含量高于乙醇提取物,泽漆中对桃蚜具有致死和拒食作用的物质可能主要存在其乳浆中。     

干旱条件下钙和赤霉素混合处理小麦种子对胚部抗氧化酶活性的影响

在20%PEG6000模拟干旱的条件下,用0.5?Cl2、50mg/L赤霉素(GA)以及0.5?Cl2 50mg/LGA浸种,研究不同浸种处理对小麦种苗胚部抗氧化酶活性的影响。结果发现,Ca GA浸种处理的种子在生长过程中,尤其是在生长的第1天和第3天,其过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)的活性在各处理中最低,抗坏血酸—过氧化物酶(APX)和谷胱甘肽还原酶(GR)的活性也维持在很低水平。表明Ca GA浸种使小麦胚部在干旱条件下保持较低的氧化状态,有利于小麦萌动和出苗。     

Multiplication of isolate R-5 of <Emphasis Type="Italic">Streptomyces galbus</Emphasis> on rhododendron leaves and its production of cell wall-degrading enzymes

 An endophytic actinomycete, isolate R-5 of Streptomyces galbus Frommer, that has promising potential as a biocontrol agent was originally isolated from field-grown rhododendron. In this study, the mode of entry of R-5 into leaves of tissue-cultured seedlings of rhododendron was investigated in connection with its production of cell wall-degrading enzymes. Light and scanning electron microscopy (SEM) revealed that R-5 grew on leaf surfaces and entered leaf tissues via stomata and that the internal mycelia grew out of stomata after colonization in host tissues. Micromanipulation at the SEM level demonstrated a prominent depression in the host surface at the interfaces with the mycelia, suggesting that such a depression could be caused by degradation of cell wall components by hydrolytic enzymes secreted from R-5 mycelia. In subsequent plate assays, R-5 produced cellulase, pectinase, xylanase, and nonspecific esterase when cultured in liquid medium. Moreover, R-5 multiplied in mineral medium containing cellulose, pectin, or xylan as a single carbon source. Thus, R-5 mycelia could degrade host cell walls at contact sites and probably utilize the degradation products as carbon sources. Received: May 16, 2002 / Accepted: July 9, 2002     

Weed Control in Wheat and Barley in the Middle East

Abstract

Wheat and barley constitute the bulk of agricultural production in the Middle East. Unfortunately, these crops are grown on marginal soils which are infested with annual and perennial weeds. The principal weed species include Avena sterilis L., Convolvulus arvensis L., and Sinapis arvensis L. and it has been estimated that yield losses due to weeds are between 20 and 30%. Control is mostly by hand pulling, hoeing or mechanical tillage, the use of herbicides being limited to about 2–5%, of the cereal area. The chemical used is usually 2,4-D but there are a few applications of barban. With the increase in wages, and unavailability of agricultural labour it is suggested that herbicide application should be widely used in the Middle East. Mixtures with benzonitriles and dicamba should be used in addition to the economical phenoxy herbicides to widen the spectrum of control of broadleaved weeds. Glyphosate may also be used after grain harvest for the control of perennial weeds and tri-allate, chlortoluron and other chemicals for the control of wild oats and other grasses. Plant breeders should study the genetic response of newly developed varieties to the most widely used herbicides.     

闹羊花素-III对菜青虫神经系统的影响

以闹羊花素 - III处理 5龄菜青虫幼虫 ,试虫乙酰胆碱酯酶 (ACh E)的活体活性和离体活性均与对照无明显差异 ,乙酰胆碱 (ACh)含量显著降低 ;对 Na - K - ATP酶、Ca2 -Mg2 - ATP酶比活力有可逆性激活作用 ,处理后 2 4 h比活力提高 ,尤以低剂量 1μg·虫 -1处理的比活力显著提高 ;处理后 6 0 h,以 1～ 3μg·虫 -1处理的比活力恢复正常 ,以 5～ 10μg·虫 -1处理的比活力显著低于对照。     

不同稻区稻瘟病菌致病型变异及基于MAS的品种对稻瘟病的抗性鉴定

稻瘟病是影响水稻生产的重要病害。监测田间稻瘟病菌菌群致病型的组成是有效推行利用抗病品种抗瘟策略的必须环节。本研究采集2015年湖北省10个稻作区的穗颈瘟标本,以单基因鉴别品系对分离得到的菌群进行致病型的测定,并分区进行品种的抗瘟性鉴定,结果表明,不同稻作区菌群致病型的组成存在差异,品种对不同稻作区菌群的抗性明显不同,单基因品系中9个抗性基因在湖北稻区受侵染程度也存在明显差异。     

危害桑树的新害虫--暗翅足距小蠹

本文综述了暗翅足距小蠹(Xylosandrus crassiusculus(Motschulsky))的研究概况,并首次记述了其可危害桑(Morus alba L.),明确了该虫为小蠹亚科(Scolytinae)江西省新纪录种。文中重点介绍了该虫的分类地位,形态特征,地理分布,寄主植物,危害特点和生物学习性,并在文章讨论部分提出了相应的防治策略。     

Carry over of bruchid,Caryedon serratus (Olivier) (Coleoptera) from field to stores through seeds of Acacia nilotica (L.) Willd. in India

Abstract

A percentage pod damage of 6.8% in Acacia nilotica (L.) Willd. by the bruchid, Caryedon serratus (Olivier) in the field was recorded by examining exit holes on the pods. The mean number of females and males that emerged from 50 pods kept in laboratory were 1.3 and 1.2 respectively. The pods of A. nilotica gathered from the field for seed purposes were found to be infested in small numbers but the bruchid population increased during storage. The infestation of seeds and carry over to stores is not only a danger to uninfested seeds of A. nilotica and other plants, important in social forestry, but the tree also acts as secondary host for the population build‐up of C. serratus which spreads to its primary host, the groundnut, Arachis hypogea (L).