Studies on viroid inducing RNA silencing from the view of plant pathology and agricultural impact

• Project/Area Number: 15580030
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Plant pathology
• Research Institution: Hirosaki University
• Principal Investigator: SANO Teruo Hirosaki University, Fac.Agric & Life Sci., Professor, 農学生命科学部, 教授 (30142699)
• Project Period (FY): 2003 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,700,000 (Direct Cost: ¥3,700,000); Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: Viroid / RNA silencing / Recovery / siRNA / Viroid homology sequence

Research Abstract

When tomato plants were infected with Potato spindle tuber viroid (PSTVd) at the cotyledon stage, primary leaf curling appeared on the 3rd leaf 14 days after inoculation. PSTVd reached the detectable all the leaves but for the meristem. PSTVd-specific siRNA also reached the detectable level in the 2nd-9th leaves. Accumulation of PSTVd and the siRNA increased to the highest level in the leaves showing severe leaf curling, however, PSTVd concentration decreased in the 9th and the upper leaves recovered from severe leaf curling. The results indicated that PSTVd-specific RNA silencing was triggered soon after PSTVd reached its detectable level, and PSTVd-specific siRNA spread quickly throughout the plant except for the meristem. The leaves being differentiated after induction of RNA silencing are to be recovered, since they are placed under the RNA silencing from the initial stage of the differentiation which resulted in the suppression of PSTVd accumulation.
Viroid-specific siRNAs of ca.21 and 23 nucleotides were detected in Hop stunt viroid-infected cucumber and hop, Hop latent viroid-infected hop and Apple fruit crinkle viroid-infected hop. On the other hand, only short siRNA of ca.21 nucleotides was detectable, in tomato infected with Cucumber mosaic virus.
Using in silica analysis by BLAST search in DNA data base, several tomato EST sequences of 18-20 nucleotides were identified. They were PCR amplified, cloned and sequenced. Using the cloned cDNAs as probes, the expression of the genes were analyzed, however, any positive signal was not identified.
Tomato seedlings obtained from healthy and viroid-infected fruits were equally infected with PSTVd challenge inoculation, which means that vertical transmission of viroid-induced RNA silencing was not apparent in tomato plants. On the other hand, the recovery was maintained in cuttings vegetatively propagated from PSTVd-infected tomato plants of the recovered stages.

Research Products

• [Journal Article] First reports of two grapevine viroids in China and the possible detection of a third (2006)
  • Author(s): Shi-fang Li, Rui Guo, Tsuji, M., Sano, T.
  • Journal Title: Plant Pathology (In Press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The first report of Coleus blumei viroid from coleus in China (2006)
  • Author(s): Shi-fang Li, Qian-fu Su, Rui Guo, Tsuji, M., Sano, T.
  • Journal Title: Plant Pathology (In Press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Double-stranded RNA-specific ribonuclease confers tolerance against Chrysanthemum stunt viroid and Tomato spotted wilt virus in transgenic chrysanthemum plants. (2005)
  • Author(s): Ogawa, T., Toguri, T., Kudoh, H., Okamura, M., Monma, T., Yoshioka, M., Kato, K., Hagiwara, Y., Sano,T.
  • Journal Title: Breeding Science 55 Pages: 49-55
  • NAID: 10014472970
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Complete nucleotide sequences of attenuated and severe isolates of Leek yellow stripe virus from garlic in northern Japan : Identification of three distinct virus types in garlic and leek world-wide. (2005)
  • Author(s): Takaki, F., Sano, T., Yamashita, K., Fujita, T., Ueda, K., Kato,T.
  • Journal Title: Arch. Virol. 150 Pages: 1135-1149
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Occurrence of Chrysanthemum chlorotic mottle viroid in Japan (2005)
  • Author(s): Yamamoto, H., Sano, T.
  • Journal Title: J. Gen. Plant Pathol. 71(2) Pages: 156-157
  • NAID: 10015507997
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Patterning of virus-infected soybean seed coat is associated with suppression of endogenous silencing of chalcone synthase genes (2005)
  • Author(s): Senda, M., Masuta, C., Ohnishi, K., Goto, K., Kasai, A., Sano, T., Hong, J.S., Macfarlane, S.
  • Journal Title: Plant Cell 16 Pages: 807-818
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Double-stranded RNA-specific ribonuclease confers tolerance against Chrysanthemum stunt viroid and Tomato spotted wilt virus in transgenic chrysanthemum plants. (2005)
  • Author(s): Ogawa, T., Toguri, T., Kudoh, H., Okamura, M., Monma, T., Yoshioka, M., Kato, K, Hagiwara, Y., Sano, T.
  • Journal Title: Breeding Science 55 Pages: 49-55
  • NAID: 10014472970
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Complete nucleotide sequences of attenuated and severe isolates of Leek yellow stripe virus from garlic in northern Japan : Identification of three distinct virus types in garlic and leek world-wide. (2005)
  • Author(s): Takaki, F., Sano, T., Yamashita, K., Fujita, T., Ueda, K., Kato, T.
  • Journal Title: Arch.Virol. 150 Pages: 1135-1149
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Occurrence of Chrysanthemum chlorotic mottle viroid in Japan (2005)
  • Author(s): Yamamoto, H., Sano, T.
  • Journal Title: J.Gen.Plant Pathol. 71(2) Pages: 156-157
  • NAID: 10015507997
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Patterning of virus-infected soybean seed coat is associated with suppression of endogenous silencing of chalcone synthase genes. (2005)
  • Author(s): Senda, M., Masuta, C., Ohnishi, K., Goto, K, Kasai, A., Sano, T., Hong, J.S., Macfarlane, S.
  • Journal Title: Plant Cell 16 Pages: 807-818
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Double-stranded RNA-specific ribonuclease confers tolerance against Chrysanthemum stunt viroid and Tomato spotted wilt virus in transgenic chrysanthemum plants. (2005)
  • Author(s): Ogawa, T., Toguri, T., Kudoh, H., Okamura, M., Momma, T., Yoshioka, M., Kato, K., Hagiwara, Y., Sano, T.
  • Journal Title: Breeding Science 55 Pages: 49-55
  • NAID: 10014472970
• [Journal Article] Characterization of a new viroid strain from hops : evidence for viroid speciation by isolation in different host specie (2004)
  • Author(s): Sano, T., Yoshida, H., Goshono, Monma, T., Kawasaki, H., Ishizaki, K.
  • Journal Title: J. Gen. Plant Pathol. 70(3) Pages: 181-187
  • NAID: 10014195262
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Accumulation of short interfering RNAs characteristic of RNA silencing precedes recovery of tomato plants from severe symptom of potato spindle tuber viroid infection (2004)
  • Author(s): Sano, T., Matsuura, Y.
  • Journal Title: J. Gen. Pl. Pathol. 70(1) Pages: 50-53
  • NAID: 10014194730
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Viroids-self-replicating non-coding RNAs (in Japanese) (2004)
  • Author(s): Sano, T.
  • Journal Title: Kagaku to Seibutsu 42(8) Pages: 508-513
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of a new viroid strain from hops : evidence for viroid speciation by isolation in different host species (2004)
  • Author(s): Sano, T., Yoshida, H., Goshono, M., Monma, T., Kawasaki, H., Ishizaki, K.
  • Journal Title: J.Gen.Pl.Pathol. 70(3) Pages: 181-187
  • NAID: 10014195262
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Accumulation of short interfering RNAs characteristic of RNA silencing precedes recovery of tomato plants from severe symptom of potato spindle tuber viroid infection (2004)
  • Author(s): Sano, T., Matsuura, Y.
  • Journal Title: J.Gen.Pl.Pathol. 70(1) Pages: 50-53
  • NAID: 10014194730
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of a new viroid strain from hops : evidence for viroid speciation by isolation in different host species (2004)
  • Author(s): Sano, T., Yoshida, H., Goshono, M., Monna, 他2名
  • Journal Title: J.Gen.Pl.Pathol. 70(3) Pages: 181-187
  • NAID: 10014195262
• [Journal Article] Patterning of virus-infected soybean seed coat is associated with suppression of endogenous silencing of chalcone synthase genes. (2004)
  • Author(s): Send, M., Masuta, C., Ohnishi, K., Goto, K,.Kasai, A., Sano, T.他2名
  • Journal Title: Plant Cell 16 Pages: 807-818
• [Journal Article] ウイロイド-自律複製するノンコーディングRNA (2004)
  • Author(s): 佐野 輝男
  • Journal Title: 化学と生物 42 Pages: 508-513
  • NAID: 10016519386
• [Journal Article] Identification of a novel structural interaction in Columnea latent viroid (2003)
  • Author(s): Owens, R.A., Sano, T., Feldstein, P.A., Hu, Y., Steger,G.
  • Journal Title: Virology 313・(2) Pages: 604-614
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of a novel structural interaction in Columnea latent viroid (2003)
  • Author(s): Owens, R.A., Sano, T., Feldstein, P.A., Hu, Y., Steger, G.
  • Journal Title: Virology 313(2) Pages: 604-614
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] First reports of two grapevine viroids in China and the possible detection of a third
  • Author(s): Shi-fang Li, Rui Guo, Tsuji, M., Sano, T.
  • Journal Title: Plant Pathology (Accepted for publication 08/Dec/15)(in Press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The first report of Coleus blumei viroid from coleus in China
  • Author(s): Shi-fang Li, Qian-fu Su, Rui Guo, Tsuji, M., Sano, T.
  • Journal Title: Plant Pathology (Accepted for publication 10/Nov/05)(in Press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The first report of Coleus blumei viroid from coleus in China.
  • Author(s): Shi-fang Li, Qian-fu Su, Rui Guo, Tsuji, M., Sano T.
  • Journal Title: Plant Pathology (In Press) (Accepted for publication 10/Nov/05)
• [Journal Article] First reports of two grapevine viroids in China and the possible detection of a third
  • Author(s): Shi-fang Li, Rui Guo, Tsuji, M., Sano, T.
  • Journal Title: Plant Pathology (In Press) (Accepted for publication 08/Dec/05)
• [Book] 植物病原アトラス (2006)
  • Author(s): 佐野 輝男(分担執筆)
  • Total Pages: 270
  • Publisher: ソフトサイエンス社
• [Book] 「研究成果報告書概要(和文)」より
  • Author(s): 該当なし
• [Publications] Owens R.A.et al.: "Identification of a novel structural interaction in Columnea latent viroid"Virology. 313(2). 604-614 (2003)
• [Publications] Sano, T., Matsuura, Y: "Accumulation of short interfering RNAs characteristic of RNA silencing precedes recovery of tomato plants from severe symptom of potato spindle tuber viroid infection"J.Gen.Plant Pathol.. 70(1). 50-53 (2004)