Plant Biotechnology Vol.30 No.4

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Contents

Original Papers

Characterization of three cinnamyl alcohol dehydrogenases from Carthamus tinctorius

Safendrri Komara Ragamustari, Naoko Shiraiwa, Takefumi Hattori, Tomoyuki Nakatsubo, Shiro Suzuki, Toshiaki Umezawa. . . . . . . 315 [PDF]

We report the isolation and characterization of cDNAs encoding cinnamyl alcohol dehydrogenase (CAD) from Carthamus tinctorius (CtCAD1, CtCAD2 and CtCAD3). In silico analysis revealed that CtCAD1 and CtCAD3 were closely related to the sinapaldehyde-specific Populus tremuloides sinapyl alcohol dehydrogenase (PtreSAD). However, unlike PtreSAD, the recombinant CtCADs did not show strong preferences for either coniferaldehyde or sinapaldehyde.

Genetic disruption of CRC 12S globulin increases seed oil content and seed yield in Arabidopsis thaliana

Yuki Fujiki, Kazumasa Kudo, Hirofumi Ono, Masumi Otsuru, Yasuyo Yamaoka, Mutsumi Akita, Ikuo Nishida. . . . . . . 327 [PDF]

To examine whether genetic suppression of seed protein increases seed oil production, we obtained T-DNA--tagged mutants for the major 12S-globulin genes in Arabidopsis, CRA1, CRB and CRC, and found elevated levels of oil in crc seeds. Furthermore, cra1, crb, and crc single-mutant plants developed substantially more branches and fruits, thereby producing more seeds than wild-type plants.

Characterization of hybrids between wild and genetically modied glyphosate-tolerant soybeans

Akihiro Kubo, Mitsuko Aono, Nobuyoshi Nakajima, Toru Nishizawa, Masanori Tamaoki, Hikaru Saji. . . . . . . 335 [PDF]

We generated and characterized various hybrids between wild and GM or non-GM soybeans in a containment greenhouse, and investigated the inheritance of the cp4 epsps transgene and its effects on the hybrids. We suggest that hybrids containing half of their genes from wild soybean and half from cultivated soybean display fitness that are intermediate between wild and cultivated soybeans, whereas BC2F2 hybrids have fitness similar to that of wild soybean. We also suggest that, despite being stably inherited, the cp4 epsps gene does not affect fitness in the absence of glyphosate treatment.

Identication of genomic factors responsible for hybrid lethality in hybrids between Nicotiana nudicaulis Watson and N. tabacum L.

Hongshuo Liu, Wataru Marubashi. . . . . . . 347 [PDF]

Hybrid lethality was expressed in seedlings from crosses between Nicotiana nudicaulis and N. tabacum at 28°C. The S subgenome in N. tabacum is responsible for the lethality occurring in hybrids between N. nudicaulis and N. tabacum. Furthermore, we concluded that the Q chromosome of the S subgenome is not responsible for the hybrid lethality observed in crosses of N. nudicaulis × N. tabacum.

A eld trial to assess the environmental biosafety of codA-transgenic Eucalyptus camaldulensis cultivation

Xiang Yu, Akira Kikuchi, Etsuko Matsunaga, Akiyoshi Kawaoka, Hiroyasu Ebinuma, Kazuo N. Watanabe. . . . . . . 357 [PDF]

The first Japanese field trial of genetically modified forest trees was conducted over a period of 4 years. Three individual lines of salt tolerance conferred on Eucalyptus camaldulensis by introducing the soil microbe-derived choline oxidase (codA) gene were planted. Environmental biosafety assessment revealed that significant differences between transformants and non-transformant were not continuous although the transient difference was found. Considering the practical use of them, evaluating their impact on the ecosystem in saline conditioned fields might be important in the future.

CAD2 deciency causes both brown midrib and gold hull and internode phenotypes in Oryza sativa L. cv. Nipponbare

Taichi Koshiba, Shinya Murakami, Takefumi Hattori, Mai Mukai, Akira Takahashi, Akio Miyao, Hirohiko Hirochika, Shiro Suzuki, Masahiro Sakamoto, Toshiaki Umezawa. . . . . . . 365 [PDF]

The cad2 (cinnamyl alcohol dehydrogenase 2) null mutant isolated from retrotransposon Tos17 insertion lines of Oryza sativa L. ssp. japonica cv. Nipponbare exhibited both brown midrib (bm) and gold hull and internode (gh) phenotypes. Lignin analysis of the mutant indicated that the brown coloration in the midribs, hulls and internodes was probably due to the accumulation of cinnamaldehyde-related structures in the lignin. In addition, the enzymatic saccharification efficiency of the mutant was increased than that of the control plants, indicating that the mutant was useful to silage purposes and biofuel production.

A novel O-methyltransferase involved in the rst methylation step of yatein biosynthesis from matairesinol in Anthriscus sylvestris

Safendrri Komara Ragamustari, Tomoyuki Nakatsubo, Takefumi Hattori, Eiichiro Ono, Yu Kitamura, Shiro Suzuki, Masaomi Yamamura, Toshiaki Umezawa. . . . . . . 375 [PDF]

Yatein is an important biosynthetic precursor of the lignan podophyllotoxin. In Anthriscus sylvestris, yatein biosynthesisis preceded by two regioselective methylations involving the intermediates: thujaplicatin and 5-O-methylthujaplicatin. We report the isolation and characterization of a cDNA encoding an O-methyltransferase involved in the first methylation step, in which thujaplicatin is converted into 5-O-methylthujaplicatin.

Overexpression of DWARF AND LESION FORMATION 1 (DLE1) causes altered activation of plant defense system in Arabidopsis thaliana

Youichi Kondou, Kosuke Noguchi, Shinsuke Kutsuna, Mika Kawashima, Arata Yoneda, Mio Ishibashi, Shu Muto, Takanari Ichikawa, Miki Nakazawa, Minami Matsui, Katsushi Manabe . . . . . . . . . . . . . . . . . . . 385 [PDF]

We analyzed the gene, DWARF AND LESION FORMATION 1 (DLE1), encoding a DUF 247 protein. Overexpression of DLE1 caused not only dwarfism but also phenotypes similar to constitutive SAR-like defense responses including simulation of lesions in leaves and a highly activated PR1 gene. Microarray analysis and subcellular localization suggested that DLE1 function in the plant defense system responding to environmental stimuli including biotic stresses in the endoplasmic reticulum.

Note

Distribution of CRa in clubroot resistance (CR) cultivars of Chinese cabbage

Daisuke Aruga, Hiroki Ueno, Hideo Matsumura, Etsuo Matsumoto, Nobuaki Hayashida. . . . . . . 393 [PDF]

Clubroot resistance (CR) is an important trait for Chinese cabbage (Brassca rapa L. ssp. pekinensis). However, CR genes in the commercial cultivar have been scarcely known at molecular level. In this work, CR of 47 Chinese cabbage cultivars were analyzed phenotypically and genotypically. CRa was found in more than 80% of high resistance cultivars, and most of these cultivars retained the CRa in heterozygote. This finding could provide important and practical information for breeders and farmers.

Laticifers in mulberry exclusively accumulate defense proteins related to biotic stresses

Sakihito Kitajima, Yoshihiro Yamamoto, Kiyoo Hirooka, Chihaya Taki, Susumu Hibino. . . . . . . 399 [PDF]

We revealed that major proteins in laticifers of mulberry were all anti-insect or anti-microbial proteins. Protein accumulation pattern was different among laticifers of lignified and unlignified organs, suggesting adaptation to different potential enemies among these organs.

Response of aliphatic glucosinolate biosynthesis to signaling molecules in MAM gene knockout mutants of Arabidopsis

Yimeng Li, Xiaoyan Tang, Masami Yokota Hirai, Xiufeng Yan. . . . . . . 403 [PDF]

In Arabidopsis aliphatic glucosinolates (AGSLs) with various side-chain length are synthesized from methionine via chain elongation cycle involving MAM1 and MAM3. In response to exogenously-applied phytohormones, MAM genes exhibited different induction patterns in wild type of Arabidopsis, and changing patterns of AGSL contents were distinctive in single knockout mutants of MAM genes compared with that in wild type, suggesting that MAM1 and MAM3 play an important role in the diversity of AGSL profiles in response to hormonal changes.

Dof transcription factors control the expression of the anaphase promoting complex/cyclosome activator CCS52A1

Naoki Takahashi, Shanshan Kong, Masaaki Umeda. . . . . . . 407 [PDF]

Endoreduplication, a modified cell cycle in which DNA replication undergoes without mitosis, is a key component of growth and development of plants. CCS52A1, an activator of anaphase promoting complex/cyclosome (APC/C), is essential for the onset and progression of endoreduplication. Here we report that DNA-binding with one finger (Dof) transcription factors directly regulate the transcription of the CCS52A1 gene. Our data imply that Dof transcription factors control plant growth through the regulation of CCS52A1 expression.

ASYMMETRIC LEAVES2 and FASCIATA2 cooperatively regulate the formation of leaf adaxial-abaxial polarity in Arabidopsis thaliana

Nanako Ishibashi, Chiyoko Machida, Yasunori Machida. . . . . . . 411 [PDF]

The establishment of leaf adaxial-abaxial polarity is important for the expansion of leaf lamina in Arabidopsis thaliana. Here we show that the ASYMMETRIC LEAVES2 (AS2) and the FASCIATA2 (FAS2) are required for the establishment of leaf adaxial-abaxial polarity. AS2 encodes a plant specific protein and FAS2 encodes a component of the Chromatin Assembly Factor-1 (CAF-1) complex, which participates in the chromatin remodeling. Results of gene expression analyses in as2 fas2 double mutants suggest that AS2 and FAS2 are positively involved in the leaf adaxialization through the repression of expression of genes for leaf abaxialization.

Functional expression of an animal type-Na+-ATPase gene from a marine red seaweed Porphyra yezoensis increases salinity tolerance in rice plants

Masanori Kishimoto, Yasuka Shimajiri, Akinobu Oshima, Akira Hase, Koji Mikami, Kazuhito Akama. . . . . . . 417 [PDF]

To characterize the properties of an animal type-Na+/K+-ATPase (PyKPA1) from the marine red alga Porphyra yezoensis, transgenic rice plants that expressed the PyKPA1 cDNA under the control of CaMV 35S RNA promoter were produced. We observed transcriptional activation of the transgene, plasma membrane-localization of the gene product, and Na+-ATPase activity in the plasma membrane fraction from transgenic rice plants. Transgenic lines showed protection from a decrease in biomass in salinity conditions, demonstrating a red algal sodium pump for conferring salinity tolerance to land plants.