Original Papers

Plant regeneration from Crotalaria spectabilis hairy roots which showed inhibited growth of root-knot nematodes (Meloidogyne hapla and M. incognita) in vitro.

Akiko Ohara, Hiroyuki Daimon, Yoji Momota, Dong Poh Chin, Masahiro Mii. . . . . . . 425 [PDF]

Crotalaria spectabilis is a leguminous species, which has been used as a green manure crop antagonistic to nematodes. In this study we report the production of transgenic plants of C. spectabilis obtained from hairy roots induced by infection with a wild strain of Agrobacterium rhizogenes. When the root-knot nematodes (Meloidogyne hapla and M. incognita) were inoculated into the hairy root, the growth inhibition of these nematodes was confirmed.

Evaluating sugarcane (Saccharum sp.) cultivars for water deficit tolerance using some key physiological markers.

Suriyan Cha-um, Sukanya Wangmoon, Chareerat Mongkolsiriwatana, Muhammad Ashraf, Chalermpol Kirdmanee. . . . . . . 431 [PDF]

Screening of some commercial sugarcane cultivars for in vitro water-deficit tolerance was carried out using key physiological markers. Based upon Ward’s multivariate cluster analyses of data for proline content, photosynthetic capacity, chlorophyll fluorescence, and growth inhibition, cultivars K84-200, K95-84, K97-32 and LK92-11 were identified as water deficit tolerant, whereas K88-92, K92-80 and UT-94-2-483 as water deficit sensitive. These observations on different cultivars’ sensitivity/tolerance were confirmed by growth and yield attributes measured in a water-deficit field trial. Thus, in vitro screening could be used effectively to screen sugarcane cultivars for drought tolerance.

Rapid evaluation of effective linear energy transfer in heavy-ion mutagenesis of Arabidopsis thaliana.

Yusuke Kazama, Liqiu Ma, Tomonari Hirano, Sumie Ohbu, Yuki Shirakawa, Shin Hatakeyama, Shuuitsu Tanaka, Tomoko Abe. . . . . . . 441 [PDF]

The effect of linear energy transfer (LET) of heavy-ion beams on mutation frequency was rapidly investigated in the M1 generation using Arabidopsis heterozygous mutant of the APG 3 gene. Among the LETs we tested (22.5-61.5 keV μm-1), 30.0 keV μm-1 or more showed two times higher mutation frequencies than 22.5 keV μm-1. LETs higher than 30.0 keV μm-1 severely affected seed productivity. Our results indicate that 30.0 keV μm-1 was optimal for obtaining Arabidopsis thaliana mutants in the M2 generation.

Overexpression of an extraplastidic β-amylase which accumulates in the radish taproot influences the starch content of Arabidopsis thaliana.

Ikuo Takahashi, Toru Kuboi, Taketomo Fujiwara, Masakazu Hara. . . . . . . 447 [PDF]

We investigated the role of the major extraplastidic β-amylase of the radish (Raphanus sativus) (RsBAMY1) in starch metabolism. During growth of the radish taproot, the amylase activity was elevated, the starch content decreased, and the soluble sugar contents increased. When the RsBAMY1 gene was overexpressed in Arabidopsis, the plants grew larger and showed lower starch contents than the wild-type plant. These results suggest that the extraplastidic β-amylase may affect the starch metabolism in some plants.

Morphological changes in Ipomoea nil using chimeric repressors of Arabidopsis TCP3 and TCP5.

Michiyuki Ono, Satomi Hiyama, Yohei Higuchi, Hiroshi Kamada, Eiji Nitasaka, Tomotsugu Koyama, Nobutaka Mitsuda, Masaru Ohme-Takagi, Kimiyo Sage-Ono. . . . . . . 457 [PDF]

By transformation with cDNAs of TCP (TEOSINTE BRANCHED1, CYCLOIDEA, and PCF)3 and TCP5 of Arabidopsis thaliana with the CRES-T system, we could successfully change flower morphology of Ipomoea nil. These chimeric repressors caused an undeveloped corolla and wavy petals. These results proved this new method of making new flower shapes especially in sympetalous flowers.

High-temperature stress susceptibility of representative japonica rice cultivars derived from Norin-22: inadequate ATP supply during seed development may lead to severe damage.

Kao-Chih She, Mitsuhiro Yaeshima, Takemasa Koumoto, Mariko Ohnuma, Tomoko Hiromasa, Mio Hirai, Takuya Matsunaga, Ryouka Tashiro, Tadamasa Sasaki, Hiroaki Kusano, Hiroaki Shimada. . . . . . . 465 [PDF]

High temperature stress during seed development leads to the reduced rice grain yield with a poor milling quality. Norin-22 had a serious weakness to the high temperature stress, which was inherited to the progenies including major rice cultivars in Japan. The defective endosperm characteristics was significantly correlated to the ATP shortage in the developing seeds. An adequate amount of ATP in immature seeds may improve the trait on the tolerance to the high temperature stress.

Pathway engineering of benzylisoquinoline alkaloid biosynthesis in transgenic California poppy cells with ectopic expression of tetrahydroberberine oxidase from Coptis japonica.

Yasutaka Matsushima, Hiromichi Minami, Kentaro Hori, Fumihiko Sato. . . . . . . 473 [PDF]

(S)-Tetrahydroberberine oxidase (THBO) is the enzyme in the last step of berberine biosynthesis. Previously purified THBO from cultured C. japonica (Cj) cells was contaminated by triosephosphate isomerase. Here, we report the re-isolation of CjTHBO cDNA and its functional characterization. Transgenic California poppy cells that ectopically expressed CjTHBO under CaMV 35S promoter showed evident accumulation of dehydrocheilanthifoline and coptisine, which were expected to be generated by the reaction of CjTHBO from pathway intermediates, i.e., cheilanthifoline and stylopine.

Short Communication

Virus-induced silencing of NtmybA1 and NtmybA2 causes incomplete cytokinesis and reduced shoot elongation in Nicotiana benthamiana.

Satoshi Araki, Yasunori Machida, Masaki Ito. . . . . . . 483 [PDF]

A group of structurally related R1R2R3-Myb proteins, such as NtmybA1 and NtmybA2 from tobacco and MYB3R1 and MYB3R4 from Arabidopsis, are known to transcriptionally activate many different G2/M phase-specific genes in plants. As was previously shown for MYB3R1 and MYB3R4, we here showed that NtmybA1 and NtmybA2 are required for completion of cytokinesis and proper shoot elongation in Nicotiana benthamiana. Our results suggested that R1R2R3-Myb transcriptional activators have conserved physiological roles across different plant species.


N-Glycan structures of glycoproteins in suspension-cultured Arabidopsis thaliana T87 cells.

Hiroyuki Kajiura, Ryoji Tanaka, Ko Kato, Yuichi Hamaguchi, Tatsuji Seki, Kazuhito Fujiyama. . . . . . . 489 [PDF]

Suspension-cultured plant cells are feasible bioreactors for the production of pharmaceutical proteins. However, little is known about the effect of suspension-culture conditions on protein N-glycosylation. Here, we studied the N-glycan structures of soluble and endogenous glycoproteins at different growth stages of suspension-cultured Arabidopsis thaliana T87 cells and concluded that N-glycans have more plant-specific sugar residues as the cultivation reaches the late or end stage of plant growth.

Evaluation of anti-herbivoral genes by Agrobacterium-mediated transient expression system.

Kei Kawazu, Naoya Wasano, Kotaro Konno, Yuko Ohashi, Atsushi Mochizuki, Ichiro Mitsuhara. . . . . . . 495 [PDF]

A high-throughput system for the evaluation of anti-herbivory genes in plant is proposed, in which genes are transiently expressed in a small area of leaf via Agrobacterium-infiltration. Two anti-herbivory genes, the cry1Ab or MLX56, were transiently expressed in the leaves of tobacco, tomato and Arabidopsis followed by inoculation of lepidopteran larvae. Potent inhibitory effects of both genes against the herbivores can be detected within 5-7 days.

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

Shoichiro Yoshimura, Masaaki Komatsu, Koichiro Kaku, Masatoshi Hori, Tomohisa Ogawa, Koji Muramoto, Tomohiko Kazama, Yukihiro Ito, Kinya Toriyama. . . . . . . 501 [PDF]

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 (cv. Tachisugata) expressing cDNA of DB1 under the control of phloem-specific promoter of rice sucrose synthase-1 gene. The transgenic plants exhibited increased resistance against brown planthopper in terms of decreased survival and fecundity.

Novel intron-containing luciferase genes for quantitative analysis of both protein and mRNA.

Hideto Hayakawa, Rieko Ogura, Kazuyuki Hiratsuka, Masashi Suzuki, Masashi Ugaki. . . . . . . 505 [PDF]

Firefly luciferase (Fluc) is used as a reporter for transient gene expression assays in plants. In this study, we constructed a series of novel Fluc genes containing intronic sequences measuring 315 bp or longer, which, during quantitative reverse-transcription PCR assays, prevented PCR-amplification of the contaminating DNA and unspliced mRNA and allowed accurate amplification of only spliced mRNA. We propose that the novel Fluc genes reported here can be used to quantitate both protein and functional mRNA levels in transient gene expression assays in plants.

Development of a model system comprising Populus as a model tree and Colletotrichum gloeosporioides as a model pathogen for studying host-pathogen interactions.

Mari Narusaka, Misato Ohtani, Taku Demura, Ryutaro Shimada, Ken Shirasu, Yoshihiro Narusaka. . . . . . . 511 [PDF]

Although hybrid Populus is currently planted worldwide for biomass production and other purposes, the hybrid often lacks the tolerance of native species to cold, drought, and disease. To investigate the susceptibility of hybrid aspen plants to the fungal pathogen Colletotrichum, which has a wide range of hosts, hybrid aspen plants were inoculated with Colletotrichum species. In the results, C. gloeosporioides strains isolated from strawberry plants could infect hybrid aspen. The experimental system using hybrid aspen and C. gloeosporioides presented here will bring a lot of fruitful information, as first model system of host tree–fungal pathogen interactions.

Development of a promoter-luciferase-based high- throughput system to monitor jasmonate-mediated defense gene expression.

Masahiro Kusama, Nobuaki Urata, Rieko Ogura, Shin-ichi Ogata, Kazuyuki Hiratsuka. . . . . . . 515 [PDF]

To develop a system suitable for JA-mediated gene expression monitoring, we compared the expression patterns of Arabidopsis gene promoters obtained from PDF1.2 and VSP1 genes in response to treatment with chemicals. As a result, we found that the use of transgenic Arabidopsis seedlings harboring the VSP1-luciferase reporter gene construct enables multiwell plates to be used for conducting high-throughput assays for the screening of chemicals.