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Zhang S.,Nanjing Agricultural University | Zhang S.,Liaoning Agricultural Vocation technical College | Chen S.,Nanjing Agricultural University | Chen F.,Nanjing Agricultural University | Jiang B.,Nanjing Agricultural University
Russian Journal of Plant Physiology | Year: 2011

The stem of the creeping Chrysanthemum morifolium Ramat. cultivar 'Yuhuajinhua' responds to a gravitational stimulus by bending. Most of the calcium ion (Ca2+) present in the stem cells was concentrated in the cell walls, intercellular space, and vacuoles, with very little in the cytoplasm. An investigation of the effects of supplying exogenous Ca2+ or the Ca2+ chelator EGTA on the creeping habit and the level of endogenous IAA showed that, for at least 60 min after gravistimulation was initiated, the level of Ca2+ present in the epidermis cell walls on the lower side of the stem was maintained at a higher level than in those on the upper side. In the endodermis cells, most of the Ca2+ was located in the intercellular space, cell walls, and throughout the vacuole, with only little within the cytoplasm. Endodermis cell Ca2+ responded rapidly (within 5 min) to gravistimulation. The level of Ca2+ continued to increase within the cytoplasm for at least 30 min into the period of gravistimulation. Exogenously applied calcium chloride (CaCl2) accentuated gravity-induced bending and the IAA concentration differed between the upper and the lower sides of the gravistimulated bent stem, whereas EGTA decreased these effects. Ca2+ appears to play an important role in the gravitropism of the 'Yuhuajinhua' creeping stem, not only by changing its distribution in the epidermis cell walls and the endodermis cytoplasm, but also by regulating IAA difference between the upper and lower sides of the creeping stem. © 2011 Pleiades Publishing, Ltd.

Wang Q.J.,Nanjing Agricultural University | Wang Q.J.,Liaoning Agricultural Vocation technical College | Gao Z.H.,Nanjing Agricultural University | Tao J.M.,Nanjing Agricultural University | And 2 more authors.
Acta Horticulturae | Year: 2012

The gene of vvicor11 isolated from cowberry is a KS type dehydrin gene, which belongs to LEA D11 or LEA II protein family. LEA (late embryogenesis-abundant protein) proteins accumulate during dehydration, low temperature, freezing, abscisic acid (ABA) treatment, and seed maturation. To better understand its expression and regulation, we isolated an 1110 bp 5' region upstream starting site of the putative vvicor11 (GenBank No. FJ429387). The sequence was identified using a genome walking strategy based on the cDNA sequence of cowberry vvicor11. Computational analysis using the PLACE and PlantCARE databases showed that the sequence included 5'-UTR Pyrimidine-rich motifs and typical promoter characterization structures, such as a TATA-box and a CAAT-box, to ensure the proper transcription initiation. There were many potential cis-acting elements involved in stress, such as ABRE, LTR, TC-rich repeats, ARE, MYB (MYC), TGACG-motif, CBFHV, and so on. The presence of these motifs could suggest that the expression of vvicor11 could be induced by ABA, low temperature, drought and high salt stress, and could be a downstream target gene of the CBF transcription factor. Transient expression of a 5'- regulatory sequence deletion series showed that the degree of 5'- regulatory sequence activation was increased the longer the sequences. One of the constructions, the CP5::GUS construct was expressed in the root, stem, leaf, petal and style of tobacco without inductive treatment. This suggested that the 5'- regulatory sequence possessed at least basal expression in various organs of a different species.

Wang Q.-J.,Nanjing Agricultural University | Wang Q.-J.,Liaoning Agricultural Vocation technical College | Xu K.-Y.,Nanjing Agricultural University | Tong Z.-G.,Nanjing Agricultural University | And 7 more authors.
Plant Cell, Tissue and Organ Culture | Year: 2010

The C-repeat binding factor/dehydration responsive element binding factor 1 (CBF/DREB1) is a critical regulator of the development of tolerance to abiotic stress in plants. Here we report the cloning of the homologous gene, VviDREB1, from a central arctic plant, the cowberry Vaccinium vitis-idaea L., using rapid amplification of cDNA ends and genome walking techniques. The cDNA and genomic DNA are 887 and 3790 bp long, respectively. Alignment and phylogenetic tree analysis suggested that VviDREB1 should be classified into the A-1 group of the DREB subfamily. The 1522-bp-long promoter contains putative cis-acting elements and transcription factor binding motifs, including elements predicted to respond to phytohormones, abiotic stress, and light. The deduced amino acid sequence of the VviDREB1 cDNA contains a predicted DNA-binding domain with a three-stranded anti-parallel β-sheet and an α-helix, similar to that of the Arabidopsis thaliana ethylene-responsive-element-binding factor AtERF1, suggesting that VviDREB1 might bind to the dehydration responsive and ethylene responsive element. The expression of VviDREB1 was mainly localized to the nuclei of onion epidermal cells. Importantly, VviDREB1 expression in the cowberry was up-regulated by cold, high-salt, and abscisic acid treatment. Our findings may provide a novel basis for further studies of the acclimation and tolerance of cowberry to abiotic stresses. © 2010 Springer Science+Business Media B.V.

Guo X.-W.,ShenYang Agricultural University | Li K.,ShenYang Agricultural University | Guo Y.-S.,ShenYang Agricultural University | Xie H.-G.,Liaoning Agricultural Vocation technical College | And 2 more authors.
Biotechnology and Biotechnological Equipment | Year: 2011

At present, replant obstacle has become an important problem in grape production, and it has seriously restricted the development of the grape industry. This study compared the soil from a replant vineyard and a newly planted vineyard where vine had been grown for 30 years and 3 years respectively. A pot experiment was conducted to investigate the changes of plant growth when grape was grown on root zone soil, sterilized root zone soil and fallow soil from the corresponding vineyards. The variation of the microbial population of the rhizosphere soil, non-rhizosphere soil and fallow soil from different vineyards was studied based on PCR-DGGE approach. The amplified bands of the dominant population were sequenced. The results showed that the plants growth was suppressed by vine replanting, and the plants grew significantly stronger after replant soil sterilization. The bacterial and fungal diversity increased as the period of the grape planting extended. The diversity of the microbial population in the rhizosphere soil was greater than that in non-rhizosphere soil. Cluster analysis showed that the microbial population structure of the rhizosphere soil had the closest association with non-rhizosphere soil after vine replanting and they were different from the population structure of fallow soil. However, the population structure of the rhizosphere soil was different from that of non-rhizosphere and fallow soil in the newly planted vineyard. Comparing to the newly planted vineyard, grape replanting caused a great change in the microbial population of the rhizosphere soil. The relative abundance of Flavobacterium sp. (DQ339585) and Bacillus sp. (AY039821) decreased, while Pedobacter sp. (AJ871084) increased in number. Omphalina farinolens (EF413029) appeared, the relative abundance of Pestalotiopsis sp. (DQ657877, DQ657875, DQ657871), Phacidium lacerum (DQ470976) and Lecythophora decumbens (AF353597) decreased, while that of the fungus Pilidium acerinum voucher (AY48709) increased. Among them, Bacillus sp., Flavobacterium sp. and Pestalotiopsis sp. in the rhizosphere soil of the replant vineyard had antagonism for pathogens, and the decrease in their relative amount reduced the ability to resist pathogens. The increasing number of Pilidium acerinum voucher might relate to a serious disease after vine replanting.

Xu K.Y.,Nanjing Agricultural University | Wang S.H.,Nanjing Agricultural University | Xi L.,Nanjing Agricultural University | Xi L.,University of British Columbia | And 7 more authors.
Genetics and Molecular Research | Year: 2010

We developed a straightforward, rapid, and inexpensive method to determine transgene copy number in tobacco.The plasmid (pSSRCopy) used for tobacco transformation contains a simple sequence repeat (SSR) locus, PT1199, which was partially deleted in the middle, a homogenous SSR locus in tobacco K326. A 168-bp segment of the cloned PT1199 was shortened to 95 bp by deleting a 73-bp internal fragment. Using a pair of SSR primers, competitive PCR was amplified from genomic DNA from transgenic tobacco harboring pSSRCopy, and the two expected bands were found. The 168-bp band (SSR-168) corresponds to endogenous PT1199 and the 95-bp band (SSR-95) comes from the integrated pSSRCopy. A single copy of a transgene can be easily distinguished from multiple copies by comparing band densities. © FUNPEC-RP.

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