Mannose-Binding Lectin from Yam (Dioscorea batatas) Tubers with Insecticidal Properties againstHelicoverpa armigera(Lepidoptera: Noctuidae)

Ohizumi, Yuki; Gaidamashvili, Mariam; Ohwada, Shyuichi; Matsuda, Kazuhiro; Kominami, Junko; Nakamura‐Tsuruta, Sachiko; Hirabayashi, Jun; Naganuma, Takako; Ogawa, Tomoya; Muramoto, Koji

doi:10.1021/jf8040269

Cited by 45 publications

(32 citation statements)

References 36 publications

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“…et al, 1993; 1998). The lectins have been extensively studied in terms of their insecticidal properties for a range of economically important pests (Ohizumi et al, 2009). Some plant lectins are stable against heating, changes in pH and digestive enzymes, and reach the intestinal tract by maintaining its structure and activity (Pusztai et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

Isolation and Biochemical Characterization of Mucus Proteins in Japanese Bunching Onion (Allium fistulosum) Green Leaves

Yamazaki

Ogawa

Muramoto

et al. 2016

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Proteins contained in the green leaf mucus of Japanese bunching onion (Allium fistulosum L.) were isolated by ammonium sulfate precipitation and separated by hydrophobic interaction chromatography. The mucus proteins had an enhanced effect on tumor necrosis factor (TNF)-α production in the RAW 264 murine macrophage cell line. Protein composition was analyzed by SDS-polyacrylamide gel electrophoresis and N-terminal amino acid sequencing.The effect on RAW 264 cells was mainly attributed to mannose-binding lectin and thaumatin-like proteins, which occupied 34% of the mucus proteins. Other major proteins contained in the mucus showed homologous amino acid sequences to those of lachrymatory factor synthase, β-1,3-glucanase, 2S albumin precursor, and pathogenesis-related protein 4. Protein composition ratios in the mucus of 13 cultivars of Japanese bunching onion were also examined.Taken together, the green leaf mucus of Japanese bunching onion may be a possible source of various bioactive proteins.

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Section: Discussionmentioning

confidence: 99%

Isolation and Biochemical Characterization of Mucus Proteins in Japanese Bunching Onion (Allium fistulosum) Green Leaves

Yamazaki

Ogawa

Muramoto

et al. 2016

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“…Approximately ϳ10 mg of TSP was separated on 12% SDS-PAGE and transferred onto PVDF membrane (0.45 mm, Millipore, USA). The membrane was reacted with anti-DB1 polyclonal antiserum at 1 : 10,000 dilution (Ohizumi et al 2009) and anti-rabbit IgG Alkaline phosphatase conjugate (Promega, USA) as secondary antibody at 1 : 5,000 dilution. Bound secondary antibody was detected using nitro blue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate (Wako Chemicals, Tokyo, Japan).…”

Section: Western Blot Analysismentioning

confidence: 99%

“…AB178475 in DDBJ was revealed to be lacking a part of 5Ј signal sequence. PCR for 5Ј RACE was carried out to obtain full length cDNA using yam tuber mRNA (Ohizumi et al 2009) and the Marathon cDNA Amplification Kit (BD Bioscience Clontech). The nucleotide sequence of full length open reading frame (ORF) of DB1 cDNA was deposited to DDBJ (accession no.…”

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Expression of gene for Dioscorea batatas tuber lectin 1 in transgenic tobacco confers resistance to green-peach aphid

Kato

Hori

Ogawa

et al. 2010

Plant Biotechnology

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Dioscorea batatas tuber lectin 1 (DB1) is a storage protein isolated from yam tuber, and is shown to be a mannose-binding lectin. It has 58% amino-acid identity to insecticidal snowdrop bulb lectin GNA. In this study, we demonstrated that Ն1 mg ml Ϫ1 DB1 in an artificial diet significantly decreased the survival and fecundity of green peach aphid, Myzus persicaeca. We produced transgenic tobacco plants expressing cDNA of DB1 under the control of Cauliflower mosaic virus 35S promoter (35S-DB1) or phloem-specific promoter of rice sucrose synthase-1 gene (RSs1-DB1), and evaluate the degree of aphid resistance in whole plant bioassays. The number of survival aphids was reduced to 60% in transgenic lines with 35S-DB1 and RSs1-DB1, which accumulated DB1 at a level of 1.8% and 0.25%, respectively, of total soluble protein. Our results indicate that DB1 can be used to enhance resistance to sap-sucking insects in transgenic crops.Key words: Aphid, Dioscorea batatas tuber lectin 1, insect resistance, transgenic tobacco.Plant Biotechnology 27, 141-145 (2010) Original PaperThis article can be found at http://www.jspcmb.jp/ Materials and methods Artificial diet bioassaysGreen peach aphid (Myzus persicae) was obtained form JT (Japan Tobacco Inc. Japan). Aphids were maintained on mature plants of Nicotiana tabacum cv. Petit Havana SR1 in a controlled environment growth chamber, 25 Ϯ 1 o C, 16L/8D. The artificial diet bioassays for aphids was carried out as described by Powell et al. (1993) using an artificial diet for aphids (Dadd and Mitter 1966). DB1 was incorporated at 2, 1 and 0.1 mg ml Ϫ1 in the artificial diet. Controls were set as treatment with feed using an artificial diet containing 0 mg ml Ϫ1 DB1 or 1 mg ml Ϫ1 BSA, and treatment with no diet (no diet controls). Ten adult apterous aphids were transferred from the host plant to glass petri-dishes. Each petri-dish was sealed with a stretched Parafilm (Pechiney Plastic Packaging Inc., USA), an 500 ml artificial diet was placed on top of the stretched Parafilm, and then the artificial diet was covered with another stretched Parafilm. The diet and Parafilm were changed daily. The bioassays were conducted in an environmental growth chamber (25 Ϯ 1°C, 16L/8D), and the number of surviving adults and newborn nymphs was counted with four sets of replication. A statistical analysis was carried out using the Excel Statistics software. Transformation of tobaccoThe nucleotide sequence for a DB1 isoform, DB1 (Leu86), under the accession no. AB178475 in DDBJ was revealed to be lacking a part of 5Ј signal sequence. PCR for 5Ј RACE was carried out to obtain full length cDNA using yam tuber mRNA (Ohizumi et al. 2009) and the Marathon cDNA Amplification Kit (BD Bioscience Clontech). The nucleotide sequence of full length open reading frame (ORF) of DB1 cDNA was deposited to DDBJ (accession no. AB513659). It encodes 172 aminoacid protein with a predicted targeting-signal sequence to endoplasmic reticulum.The cDNA covering full-length ORF was PCR cloned into pGEM T-vectors (P...

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“…The insecticidal properties of DB1 have been reported against moth larvae (Helicoverpa armigera). The rate of adults emerging from pupae was reduced to 33%, when fed on 0.01% (w/v) DB1 in an artificial diet (Ohizumi et al 2009). We also demonstrated that ≥1 mg/ml DB1 in an artificial diet significantly decreased the survival and fecundity of green peach aphid, Myzus persicaeca (Kato et al 2010).…”

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Production of transgenic rice plants expressing Dioscorea batatas tuber lectin 1 to confer resistance against brown planthopper

Yoshimura¹,

Komatsu²,

Kaku³

et al. 2012

Plant Biotechnology

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Dioscorea batatas tuber lectin 1 (DB1) is a storage protein isolated from yam tuber and has been shown to be a mannose-binding lectin. Here we produced transgenic rice plants expressing cDNA of DB1 under the control of phloemspecific promoter of rice sucrose synthase-1 gene. DB1 accumulated at a level of 0.07% of total soluble protein. We then evaluated its efficacy for brown planthopper (BPH). After releasing the first instar of BPH on the transgenic rice plants, the number of survived BPH adults was reduced up to 30% compared to that of wild-type rice. The number of the next generation BPH was suppressed to 22% on average in the seven most-resistant plants compared to that of wild-type rice plants when female adult BPH was inoculated. These results demonstrated that DB1 is effective to confer BPH resistance in terms of decreased survival and fecundity.Key words: Dioscorea batatas tuber lectin 1, insect resistance, planthopper, transgenic rice.Three major sap-sucking pests of rice in Japan: brown planthopper (Nilaparvata lugens, BPH), whitebacked planthopper (Sogatella furcifera, WBPH) and small brown planthopper (Laodelphax striatellus) are known to cause severe damage to rice by sucking or acting as vectors for major viral disease. Recently, these pests have developed resistance to most commercial chemical pesticides (Matsumura et al. 2008). A transgenic approach has been explored to confer rice plants with resistance to such sap-sucking insects. One of the promising genes is encoding mannose-binding protein such as snowdrop lectin (Galanthus nivalis agglutinin; GNA) and garlic leaf lectin (Allium sativum agglutinin from leaf; ASAL). Expression of GNA or ASAL in rice plants has been shown to confer substantial resistance to BPH, WBPH, and green rice planthopper in terms of increased insect mortality, retarded development and decreased fecundity (Rao et al. 1998;Nagadhara et al. 2003Nagadhara et al. , 2004Saha et al. 2006a; Yarashi et al. 2008). Mannose-binding proteins, in general, are known to bind to mannose-containing glycoproteins in the mid-guts of insects, and inhibit growth and development (Sauvion et al. 1996).We have been focusing on Dioscorea batatas tuber lectin 1 (DB1), which has been isolated from yam tuber, Dioscorea batatas Decne., as a storage protein. DB1 is considered to be least non-harmful to human beings, as we Japanese eat raw tubers without boiling them. DB1 is a mannose-binding lectin (23 kDa) consisting of identical 12-kDa subunits. It has 58% and 48% amino-acid identity to GNA and ASAL, respectively, and is classified in the GNA-related lectin family (Gaidamashvili et al. 2004). The insecticidal properties of DB1 have been reported against moth larvae (Helicoverpa armigera). The rate of adults emerging from pupae was reduced to 33%, when fed on 0.01% (w/v) DB1 in an artificial diet (Ohizumi et al. 2009). We also demonstrated that ≥1 mg/ml DB1 in an artificial diet significantly decreased the survival and fecundity of green peach aphid, Myzus persicaeca (Kato et al. 2010). Th...

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Mannose-Binding Lectin from Yam (Dioscorea batatas) Tubers with Insecticidal Properties againstHelicoverpa armigera(Lepidoptera: Noctuidae)

Cited by 45 publications

References 36 publications

Isolation and Biochemical Characterization of Mucus Proteins in Japanese Bunching Onion (<i>Allium fistulosum</i>) Green Leaves

Isolation and Biochemical Characterization of Mucus Proteins in Japanese Bunching Onion (<i>Allium fistulosum</i>) Green Leaves

Expression of gene for Dioscorea batatas tuber lectin 1 in transgenic tobacco confers resistance to green-peach aphid

Production of transgenic rice plants expressing Dioscorea batatas tuber lectin 1 to confer resistance against brown planthopper

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