Xianwen Zhang

Expression analysis of nine rice heat shock protein genes under abiotic stresses and ABA treatment.

Expression profiles of nine rice heat shock protein genes (OsHSPs) were analyzed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The nine genes exhibited distinctive expression in different organs. Expression of nine OsHSP genes was affected differentially by abiotic stresses and abscisic acid (ABA). All nine OsHSP genes were induced strongly by heat shock treatment, whereas none of them were induced by cold. The transcripts of OsHSP80.2, OsHSP71.1 and OsHSP23.7 were increased during salt tress treatment. Expression of OsHSP80.2 and OsHSP24.1 genes were enhanced while treated with 10% PEG. Only OsHSP71.1 was induced by ABA while OsHSP24.1 was suppressed by ABA. These observations imply that the nine OsHSP genes may play different roles in plant development and abiotic stress responses.

Expression Profile in Rice Panicle: Insights into Heat Response Mechanism at Reproductive Stage

Rice at reproductive stage is more sensitive to environmental changes, and little is known about the mechanism of heat response in rice panicle. Here, using rice microarray, we provided a time course gene expression profile of rice panicle at anther developmental stage 8 after 40°C treatment for 0 min, 20 min, 60 min, 2 h, 4 h, and 8 h. The identified differentially expressed genes were mainly involved in transcriptional regulation, transport, cellular homeostasis, and stress response. The predominant transcription factor gene families responsive to heat stress were Hsf, NAC, AP2/ERF, WRKY, MYB, and C2H2. KMC analysis discovered the time-dependent gene expression pattern under heat stress. The motif co-occurrence analysis on the promoters of genes from an early up-regulated cluster showed the important roles of GCC box, HSE, ABRE, and CE3 in response to heat stress. The regulation model central to ROS combined with transcriptome and ROS quantification data in rice panicle indicated the great importance to maintain ROS balance and the existence of wide cross-talk in heat response. The present study increased our understanding of the heat response in rice panicle and provided good candidate genes for crop improvement.

Transcriptome profile reveals heat response mechanism at molecular and metabolic levels in rice flag leaf

Flag leaf is one of the key photosynthesis organs during rice reproductive stage. A time course microarray analysis of rice flag leaf was done after 40°C treatment for 0 min, 20 min, 60 min, 2h, 4h, and 8h. The identified significant heat responsive genes were mainly involved in transcriptional regulation, transport, protein binding, antioxidant, and stress response. KMC analysis discovered the time-dependent gene expression pattern under heat. MapMan analysis demonstrated that, under heat treatment, Hsp genes and genes involved in glycolysis and ubiquitin-proteasome were enhanced, and genes involved in TCA, carotenoid, dihydroflavonol and anthocyanin metabolisms and light-reaction in the photosynthesis were widely repressed. Meanwhile, some rate-limiting enzyme genes in shikimate, lignin, and mevalonic acid metabolisms were up-regulated, revealing the importance of maintaining specific secondary metabolites under heat stress. The present study increased our understanding of heat response in rice flag leaf and provided good candidate genes for crop improvement.

Expression and Promoter Analysis of Six Heat Stress-Inducible Genes in Rice

During the long evolutionary process, plant gradually formed a series of strategies and mechanisms to cope with stress environment such as drought, heat, cold, and high salinity. Six highly heat responsive genes were identified in rice by microarray data analysis. The qRT-PCR analysis confirmed that the expression of these six genes were highly heat inducible and moderately responded to salt stress, polyethylene glycol, and abscisic acid treatment, but little affected by cold treatment. Promoters of the three highly heat-inducible genes (OsHsfB2cp, PM19p, and Hsp90p) were used to drive GUS gene expression in rice. The results of the GUS gene expression, histochemical staining, and GUS activities in panicles and flag leaves of the transgenic rice plants confirmed high heat-induced GUS activities and moderate drought-induced activities. The three promoters exhibited similar high activity lever in rice leaf under heat, but OsHsfB2cp and PM19p showed much higher activities in panicles under heat stress. Our work confirmed that the OsHsfB2c and PM19 promoters were highly heat inducible and further characterization and reconstruction of cis-elements in their promoters could lead to the development of highly effective heat-inducible promoters for plant genetic engineering.

Heat shock factor OsHsfB2b negatively regulates drought and salt tolerance in rice

Key messageExpression ofOsHsfB2bwas strongly induced by heat, salt, ABA and PEG treatments. Drought and salt tolerances were significantly decreased byOsHsfB2boverexpression, but were enhanced by RNA interference.AbstractPlants have more than 20 heat shock factors (Hsfs) that were designated class A, B, and C. Many members of Class A Hsfs were characterized as activators of transcription, but the functional roles of class B and C Hsfs have not been fully recognized. OsHsfB2b is a member of class B Hsfs in rice (Oryza sativa). Expression of OsHsfB2b was strongly induced by heat, salt, abscisic acid (ABA) and polyethylene glycol (PEG) treatments but was almost not affected by cold stress. Drought and salt tolerances were significantly decreased in OsHsfB2b-overexpressing transgenic rice, but were enhanced in the OsHsfB2b-RNAi transgenic rice. Under drought stress, the OsHsfB2b-overexpressing transgenic rice exhibited increased relative electrical conductivity (REC) and content of malondialdehyde (MDA) and decreased proline content compared with the wild type, while the lower REC and MDA content and increased proline content were found in the OsHsfB2b-RNAi transgenic rice. These results suggest that OsHsfB2b functions as a negative regulator in response to drought and salt stresses in rice, with its existing B3 repression domain (BRD) that might be necessary for the repressive activity. The present study revealed the potential value of OsHsfB2b in genetic improvement of rice.

Characterization of the global transcriptome for cotton (Gossypium hirsutum L.) anther and development of SSR marker

Cotton is an important fiber plant, and its attractive to elucidate the molecular mechanism of anther development due to the close relationship between the anther fertility and boll-setting, and also fiber yield. In the present paper, 47.2 million paired-end reads with average length of 82.87 bp from the anthers of TM-1 (Gossypium hirsutum L.), a genetic standard line, were generated through transcriptome sequencing, and 210,965 unigenes of more than 100 bp were obtained. BLAST, KEGG, COG, and GO analyses showed that the genes were enriched in the processes of transcription, translation, and post-translation as well as hormone signal transduction, the transcription factor families, and cell wall-related genes mainly participating in cell expansion and carbohydrate metabolism. Further analysis identified 11,153 potential SSRs. A suit of 5122 primer pair sequences were designed, and 82 of 300 randomly selected primer pairs produced reproducible amplicons that were polymorphic among 22 cotton accessions from G. hirsutum, Gossypium barbadense and Gossypium arboreum. The UPGMA clustering analysis further confirmed high quality and effectiveness of these novel SSR markers. The present study provided insights into the transcriptome profile of the cotton and established a public information platform for functional genomics and molecular breeding.

OsGL1-3 is involved in cuticular wax biosynthesis and tolerance to water deficit in rice.

Cuticular wax covers aerial organs of plants and functions as the outermost barrier against non-stomatal water loss. We reported here the functional characterization of the Glossy1(GL1)-homologous gene OsGL1-3 in rice using overexpression and RNAi transgenic rice plants. OsGL1-3 gene was ubiquitously expressed at different level in rice plants except root and its expression was up-regulated under ABA and PEG treatments. The transient expression of OsGL1-3–GFP fusion protein indicated that OsGL1-3 is mainly localized in the plasma membrane. Compared to the wild type, overexpression rice plants exhibited stunted growth, more wax crystallization on leaf surface, and significantly increased total cuticular wax load due to the prominent changes of C30–C32 aldehydes and C30 primary alcohols. While the RNAi knockdown mutant of OsGL1-3 exhibited no significant difference in plant height, but less wax crystallization and decreased total cuticular wax accumulation on leaf surface. All these evidences, together with the effects of OsGL1-3 on the expression of some wax synthesis related genes, suggest that OsGL1-3 is involved in cuticular wax biosynthesis. Overexpression of OsGL1-3 decreased chlorophyll leaching and water loss rate whereas increased tolerance to water deficit at both seedling and late-tillering stages, suggesting an important role of OsGL1-3 in drought tolerance.

Microarray Data Uncover the Genome-wide Gene Expression Patterns in Response to Heat Stress in Rice Post-meiosis Panicle

To comprehend the gene expression profile in rice panicle under high temperature, Agilent 4×44K rice oligo microarray experiments were carried out using rice post-meiosis panicle treated at 40 centigrade degree for 0 min, 10 min, 20 min, 60 min, and 2 h. The time course differentially expressed genes under heat stress were mainly involved in protein binding, catalysis, stress response, and cellular process. The significantly changed genes during heat treatment were mainly up-regulated. Among heat-responsive (HR) genes, the predominant transcription factor gene families were Hsf, NAC, AP2/ERF, WRKY, MYB, and C2H2. Fifty four of the HR genes were functionally characterized based on OGRO database, and most of these characterized HR genes were related to stress resistance, panicle flower and sterility. The MapMan analysis demonstrated that, under heat stress, the HR genes were enriched in the pathways related to biotic and abiotic stress, cell cycle, development, ubiquitinproteasome system, lipid and secondary metabolisms. Comparative transcriptome analysis identified 113 potential anther meiosis-related target genes under heat stress. These data revealed the great importance of multiple functions of candidate genes and protein homeostasis in response to heat stress in rice panicle at post-meiosis stage.

The Arabidopsis AtUNC-93 acts as a positive regulator of abiotic stress tolerance and plant growth via modulation of ABA signaling and K+ homeostasis

Potassium (K+) is one of the essential macronutrients required for plant growth and development, and the maintenance of cellular K+ homeostasis is important for plants to adapt to abiotic stresses and growth. However, the mechanism involved has not been understood clearly. In this study, we demonstrated that AtUNC-93 plays a crucial role in this process under the control of abscisic acid (ABA). AtUNC-93 was localized to the plasma membrane and mainly expressed in the vascular tissues in Arabidopsis thaliana. The atunc-93 mutants showed typical K+-deficient symptoms under low-K+ conditions. The K+ contents of atunc-93 mutants were significantly reduced in shoots but not in roots under either low-K+ or normal conditions compared with wild type plants, whereas the AtUNC-93-overexpressing lines still maintained relatively higher K+ contents in shoots under low-K+ conditions, suggesting that AtUNC-93 positively regulates K+ translocation from roots to shoots. The atunc-93 plants exhibited dwarf phenotypes due to reduced cell expansion, while AtUNC-93-overexpressing plants had larger bodies because of increased cell expansion. After abiotic stress and ABA treatments, the atunc-93 mutants was more sensitive to salt, drought, osmotic, heat stress and ABA than wild type plants, while the AtUNC-93-overexpressing lines showed enhanced tolerance to these stresses and insensitive phenotype to ABA. Furthermore, alterations in the AtUNC-93 expression changed expression of many ABA-responsive and stress-related genes. Our findings reveal that AtUNC-93 functions as a positive regulator of abiotic stress tolerance and plant growth by maintaining K+ homeostasis through ABA signaling pathway in Arabidopsis.

Arabidopsis TEMPRANILLO1 transcription factor AtTEM1 negatively regulates drought tolerance

Arabidopsis TEMPRANILLO 1 (AtTEM1) is a member of the RAV transcription factor subfamily and has plant-specific AP2 and B3 domains. This study demonstrated that AtTEM1 is localized in nuclei. Expression of AtTEM1 was discovered to be high in the flower, stem, rosette, and cauline leaf, but low in the silique and root tissue, and was strongly induced by drought and mannitol treatments. Compared with wild-type plants, the 35S::AtTEM1 plants bolted later and produced more rosette leaves before flowering, whereas, the AtTEM1 mutant plants (tem1) bolted earlier but produced an almost equal number of rosette leaves before flowering. Drought tolerance was significantly reduced in the 35S::AtTEM1 transgenic Arabidopsis plants, but was enhanced in the tem1 plants. Under drought and mannitol stresses, the 35S::AtTEM1 transgenic Arabidopsis exhibited increased relative electrical conductivity (REC), increased malondialdehyde (MDA) content, and decreased proline content compared with the wild type, whereas lower REC and MDA content and higher proline content were found in the tem1 plants. These results suggest that AtTEM1 in Arabidopsis functions as a negative regulator in response to drought and mannitol stresses. Therefore, AtTEM1 transcription factor is proposed to be a versatile regulator reacting negatively to both flowering time and drought stress.