A Brassica napus transcript encoding a protein related to the Kunitz protease inhibitor family accumulates upon water stress in leaves, not in seeds.

Downing, WL; Mauxion, Fabienne; Fauvarque, MO; Reviron, MP; Vienne, Dominique de; Vartanian, Nicole; Giraudat, Jérôme

doi:10.1046/j.1365-313x.1992.t01-11-00999.x

Cited by 184 publications

(84 citation statements)

References 16 publications

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“…To determine GH3.9 expression in different genotypes and RNAi transgenic lines, total RNA was isolated from tissue comprising both mature and immature siliques and some flowers using an SDS-phenol extraction procedure followed by LiCl precipitation essentially as described (Downing et al 1992). Total RNA (15 μg) was electrophoresed on a 1.2% formaldehyde-agarose gel and blotted overnight in 20X SSC (3 M sodium chloride, 0.3 M sodium citrate, pH 7.0) to a Hybond-N membrane (Amersham Biosciences, Piscataway, NJ, USA).…”

Section: Semi-quantitative Rt-pcr and Northern (Rna) Blot Analysesmentioning

confidence: 99%

Arabidopsis thaliana GH3.9 influences primary root growth

Khan

Stone

2007

Planta

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Auxins regulate a complex signal transduction network to direct plant development. Auxin-responsive genes fit into three major classes: the so-called auxin/indole-3-acetic acid (Aux/IAA), the GH3, and the small auxinup RNA (SAUR) gene families. The 20-member Arabidopsis thaliana GH3 gene family has been subdivided into three groups. In vitro studies have shown that most Group II members function as IAA-amido synthetases to conjugate amino acids to the plant hormone auxin. Here we report the role of a previously uncharacterized GH3 gene family member, GH3.9, in root growth. Unlike most other Group II family members, GH3.9 expression was repressed by low concentrations of exogenous IAA in seedlings. Transgenic plants harboring a GH3.9 promoter::reporter gene construct indicate that GH3.9 is expressed in the root-hypocotyl junction, leaves and the shoot apical meristem of young seedlings, in mature embryos, and in the root vascular tissue. Expression was also observed in lateral root tips when seedlings were treated with exogenous IAA. Inverse PCR was used to identify an activation tagged T-DNA insertion in chromosome 2 near the 5′UTR region of At2g47750 (GH3.9). Plants homozygous for the T-DNA insertion (gh3.9-1 mutants) had reduced GH3.9 expression, no obvious effects on apical dominance or leaf morphology, greater primary root length, and increased sensitivity to indole-3-acetic acid (IAA)-mediated root growth inhibition. Additional T-DNA insertion alleles and transgenic plants with reduced GH3.9 transcript levels due to RNA-interference (RNAi) also showed these same phenotypes. Our results provide new information on the function of GH3.9 in roots where it is likely to control auxin activity through amino acid conjugation.

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Section: Semi-quantitative Rt-pcr and Northern (Rna) Blot Analysesmentioning

confidence: 99%

Arabidopsis thaliana GH3.9 influences primary root growth

Khan

Stone

2007

Planta

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“…Embryos of different developmental stages were separated from siliques and RNA was isolated according to Parcy et al (1994) and Downing et al (1992) …”

Section: Rt-pcr Analysis Of Gene Expressionmentioning

confidence: 99%

The Peroxisome Deficient Arabidopsis Mutant sse1 Exhibits Impaired Fatty Acid Synthesis

2004

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The Arabidopsis Shrunken Seed 1 (SSE1) gene encodes a homolog of the peroxisome biogenesis factor Pex16p, and a loss-of-function mutation in this gene alters seed storage composition. Two lines of evidence support a function for SSE1 in peroxisome biogenesis: the peroxisomal localization of a green fluorescent protein-SSE1 fusion protein and the lack of normal peroxisomes in sse1 mutant embryos. The green fluorescent protein-SSE1 colocalizes with the red fluorescent protein (RFP)-labeled peroxisomal markers RFP-peroxisome targeting signal 1 and peroxisome targeting signal 2-RFP in transgenic Arabidopsis. Each peroxisomal marker exhibits a normal punctate peroxisomal distribution in the wild type but not the sse1 mutant embryos. Further studies reported here were designed toward understanding carbon metabolism in the sse1 mutant. A time course study of dissected embryos revealed a dramatic rate decrease in oil accumulation and an increase in starch accumulation. Introduction of starch synthesis mutations into the sse1 background did not restore oil biosynthesis. This finding demonstrated that reduction in oil content in sse1 is not caused by increased carbon flow to starch. To identify the blocked steps in the sse1 oil deposition pathway, developing sse1 seeds were supplied radiolabeled oil synthesis precursors. The ability of sse1 to incorporate oleic acid, but not pyruvate or acetate, into triacylglycerol indicated a defect in the fatty acid biosynthetic pathway in this mutant. Taken together, the results point to a possible role for peroxisomes in the net synthesis of fatty acids in addition to their established function in lipid catabolism. Other possible interpretations of the results are discussed.

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“…Seed RNA was extracted following Downing (1992). RNA from other plant tissues was isolated as described in Procissi et al (2003).…”

Section: Northern-blot Analysismentioning

confidence: 99%

The Putative Arabidopsis Homolog of Yeast Vps52p Is Required for Pollen Tube Elongation, Localizes to Golgi, and Might Be Involved in Vesicle Trafficking

et al. 2004

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The screening of the Versailles collection of Arabidopsis T-DNA transformants allowed us to identify several male gametophytic mutants, including poky pollen tube (pok). The pok mutant, which could only be isolated as a hemizygous line, exhibits very short pollen tubes, explaining the male-specific transmission defect observed in this line. We show that the POK gene is duplicated in the Arabidopsis genome and that the predicted POK protein sequence is highly conserved from lower to higher eukaryotes. The putative POK homolog in yeast (Saccharomyces cerevisiae), referred to as Vps52p/SAC2, has been shown to be located at the late Golgi and to function in a complex with other proteins, Vps53p, Vps54p, and Vps51p. This complex is involved in retrograde trafficking of vesicles between the early endosomal compartment and the trans-Golgi network. We present the expression patterns of the POK gene and its duplicate P2 in Arabidopsis, and of the putative Arabidopsis homologs of VPS53 and VPS54 of yeast. We show that a POK::GFP fusion protein localizes to Golgi in plant cells, supporting the possibility of a conserved function for Vps52p and POK proteins. These results, together with the expression pattern of the POK::GUS fusion and the lack of plants homozygous for the pok mutation, suggest a more general role for POK in polar growth beyond the pollen tube elongation process.Pollen tube growth is a vital process during male gametophyte development, since it allows male gametes to reach the ovules and achieve fertilization (Preuss, 1995). The elongation of the pollen tube, as well as that of animal axons, plant root hairs, fungal hyphae, and moss protonema, is achieved by a polarized mode of growth, termed tip growth (FranklinTong, 1999;Palanivelu and Preuss, 2000;Hepler et al., 2001), which involves the tip-localized exocytosis of Golgi-derived vesicles containing cell wall precursors (Franklin-Tong, 1999). The tip of the pollen tube is devoid of organelles, but contains almost exclusively Golgi-derived vesicles (Geitmann and Emons, 2000). In the shank of the tubes, an inverse fountain pattern of cytoplasmic streaming is observed; organelles and vesicles are transported toward the apex in the cortical region and basipetally in the central cytoplasm (Pierson et al., 1990). Actin filaments and microtubules, which are organized in longitudinal arrays more or less parallel to the elongation axis, act as tracks for cytoplasmic streaming and allow delivery of vesicles to the tip (Pierson et al., 1990;Vidali and Hepler, 2001). The use of membrane structure dyes, such as FM1-43 or FM4-64 (Parton et al., 2001;Camacho and Malhó , 2003), has revealed dynamic endo/ exocytosis processes at the tip of the pollen tube, but the molecular events underlying these processes are still poorly understood. The recent identification of the Golgi-associated tobacco (Nicotiana tabacum), Rab GTPase NtRab2, predominantly expressed in pollen, suggests that tip growth and vesicle trafficking could be tightly linked (Cheung et al., 2002). Moreover, a...

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A Brassica napus transcript encoding a protein related to the Kunitz protease inhibitor family accumulates upon water stress in leaves, not in seeds.

Cited by 184 publications

References 16 publications

Arabidopsis thaliana GH3.9 influences primary root growth

Arabidopsis thaliana GH3.9 influences primary root growth

The Peroxisome Deficient Arabidopsis Mutant sse1 Exhibits Impaired Fatty Acid Synthesis

The Putative Arabidopsis Homolog of Yeast Vps52p Is Required for Pollen Tube Elongation, Localizes to Golgi, and Might Be Involved in Vesicle Trafficking

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