Two New Group 3 LEA Genes of Wheat and Their Functional Analysis in Yeast

The group 3 late embryogenesis abundant （LEA） proteins are thought to protect cells from stresses associated with dehydration during periods of water deficit. To investigate the functions of different members of the group 3 LEA genes, we isolated and characterized two new group 3 LEA genes, namely TaLEA2 and TaLEA3, from wheat （Triticum aestivum L.） and introduced TaLEA2 and TaLEA3 into Saccharmyces cerevisiae to examine the effect of these genes on yeast cell tolerance to osmotic, salt, and cold stresses. The TaLEA2 gene encoded a protein of 211 amino acids and possessed five repeats of 11-mer amino acid motifs. The TaLEA3 gene encoded a polypeptide of 211 amino acids with nine repeated units. Overexpression of TaLEA2 and TaLEA3 improved stress tolerance in transgenic yeast cells when cultured in medium containing sorbitol, salt and-20℃ freezing treatments respectively. However, the yeast transformants with TaLEA2 seemed to be more tolerant to hyperosmotic and freezing stress than transformants with TaLEA3. This implies that a close relationship exists between function and the number of repeats of the 11- mer amino acid motif in the group 3 LEA protein.     

Plant Functional Types Differ in Their Long-term Nutrient Response to eCO2 in an Extensive Grassland

Ecosystems - Increasing atmospheric CO2 enhances plant biomass production and may thereby change nutrient concentrations in plant tissues. The objective of this study was to identify the effect of...     

Functional and Evolutionary Characterization of the CONSTANS Gene Family in Short-Day Photoperiodic Flowering in Soybean

CONSTANS (CO) plays a central role in photoperiodic flowering control of plants. However, much remains unknown about the function of the CO gene family in soybean and the molecular mechanisms underlying short-day photoperiodic flowering of soybean. We identified 26 CO homologs (GmCOLs) in the soybean genome, many of them previously unreported. Phylogenic analysis classified GmCOLs into three clades conserved among flowering plants. Two homeologous pairs in Clade I, GmCOL1a/GmCOL1b and GmCOL2a/GmCOL2b, showed the highest sequence similarity to Arabidopsis CO. The mRNA abundance of GmCOL1a and GmCOL1b exhibited a strong diurnal rhythm under flowering-inductive short days and peaked at dawn, which coincided with the rise of GmFT5a expression. In contrast, the mRNA abundance of GmCOL2a and GmCOL2b was extremely low. Our transgenic study demonstrated that GmCOL1a, GmCOL1b, GmCOL2a and GmCOL2b fully complemented the late flowering effect of the co-1 mutant in Arabidopsis. Together, these results indicate that GmCOL1a and GmCOL1b are potential inducers of flowering in soybean. Our data also indicate rapid regulatory divergence between GmCOL1a/GmCOL1b and GmCOL2a/GmCOL2b but conservation of their protein function. Dynamic evolution of GmCOL regulatory mechanisms may underlie the evolution of photoperiodic signaling in soybean.     

Identification of 14-3-3 Family in Common Bean and Their Response to Abiotic Stress

14-3-3s are a class of conserved regulatory proteins ubiquitously found in eukaryotes, which play important roles in a variety of cellular processes including response to diverse stresses. Although much has been learned about 14-3-3s in several plant species, it remains unknown in common bean. In this study, 9 common bean 14-3-3s (PvGF14s) were identified by exhaustive data mining against the publicly available common bean genomic database. A phylogenetic analysis revealed that each predicted PvGF14 was clustered with two GmSGF14 paralogs from soybean. Both epsilon-like and non-epsilon classes of PvGF14s were found in common bean, and the PvGF14s belonging to each class exhibited similar gene structure. Among 9 PvGF14s, only 8 are transcribed in common bean. Expression patterns of PvGF14s varied depending on tissue type, developmental stage and exposure of plants to stress. A protein-protein interaction study revealed that PvGF14a forms dimer with itself and with other PvGF14 isoforms. This study provides a first comprehensive look at common bean 14-3-3 proteins, a family of proteins with diverse functions in many cellular processes, especially in response to stresses.     

小麦PAL基因的克隆及赤霉菌诱导下的表达分析

利用苯丙氨酸解氨酶(PAL,phenylalanine ammonia-lyase)基因保守区域从小麦抗赤霉病材料苏麦3号中克隆获得4个PAL基因,分别命名为Ta PAL1、Ta PAL2、Ta PAL3、Ta PAL4。4个基因的开放阅读框(ORF,open reading frame)长度分别为2142 bp、2016 bp、2118 bp和2139 bp,分别编码714个、672个、706个和713个氨基酸。基因序列比对发现其相似性达到88.35%,所编码的氨基酸相似性为91.92%,氨基酸序列分析表明4个基因都包含HAL-PAL结构域及PAL结构域。通过接种禾谷镰刀菌,利用荧光定量PCR对PAL基因进行表达分析发现,4个PAL基因全部为上调表达,其中Ta PAL2、Ta PAL3和Ta PAL4最为明显。PAL基因的上调表达,说明PAL基因在小麦抵抗赤霉病菌侵染的机制中可能起着重要作用。     

Rtm1: A Member of a New Family of Telomeric Repeated Genes in Yeast

We have isolated a new yeast gene called RTM1 whose overexpression confers resistance to the toxicity of molasses. The RTM1 gene encodes a hydrophobic 34-kD protein that contains seven potential transmembrane-spanning segments. Analysis of a series of industrial strains shows that the sequence is present in multiple copies and in variable locations in the genome. RTM loci are always physically associated with SUC telomeric loci. The SUC-RTM sequences are located between X and Y' subtelomeric sequences at chromosome ends. Surprisingly RTM sequences are not detected in the laboratory strain X2180. The lack of this sequence is associated with the absence of any SUC telomeric gene previously described. This observation raises the question of the origin of this nonessential gene. The particular subtelomeric position might explain the SUC-RTM sequence amplification observed in the genome of yeasts used in industrial biomass or ethanol production with molasses as substrate. This SUC-RTM sequence dispersion seems to be a good example of genomic rearrangement playing a role in evolution and environmental adaptation in these industrial yeasts.     

A Transient Transformation System for the Functional Characterization of Genes Involved in Stress Response

In this study, we developed a simple and efficient transient transformation system, which can be used in homologous expression or reverse genetic study of the plants. A system for characterizing gene function in response to stress tolerance was also developed based on this transformation method. The overexpression and RNAi-silencing of a bZIP gene from Tamarix hispida, ThbZIP1, were performed in T. hispida using this transformation method. Real-time PCR showed that the expression of ThbZIP1 was highly up- and down-regulated in the plants with overexpression and RNAi-silenced expression of ThbZIP1, respectively, when compared with control plants (transiently transformed with empty pROK2). A physiological study showed that ThbZIP1 can enhance the activities of both peroxidase (POD) and superoxide dismutase (SOD), and decrease electrolyte leakage rate and levels of reactive oxygen species (ROS) and malondialdehyde (MDA) under salt stress conditions. Furthermore, ThbZIP1 is found to mediate stress tolerance by regulating the expression of SOD and POD genes. These results suggested that this transient transformation system is an effective method for determining the function of a gene in response to abiotic stress in plants.     

Structural and Functional Characterization of Human Chromosome 13q14 and Its Potential Tumor Suppressor Genes

Works on chromosome 13 mapping supported by the Russian program Human Genome are reviewed. Emphasis is placed on studies of region 13q14.3, which is often lost in some human tumors and potentially contains tumor suppressor genes (TSG). A strategy of TSG search is described. As the resolution of genome analysis improved, a minimal overlap of genetic loss in B-cell chronic lymphocytic leukemia (B-CLL) was established for chromosome 13. A map of expressed sequences was constructed for the region containing the overlap, and candidate TSG of chromosome 13q14 were identified. The candidate genes were analyzed both structurally and functionally, and their possible role in tumorigenesis was considered. Assuming haploinsufficiency as a genetic mechanism controlling B-CLL, a new strategy was proposed for mutation screening aimed at identifying potential TSG of region 13q14.     

Cloning and Characterization of Novel CIS Family Genes

We have reported two JAK-signaling modulators, CIS (cytokine-inducible SH2 protein) and JAB (JAK2 binding protein), which are structurally related. Here we cloned three additional CIS family genes (CIS2, CIS3, and CIS4) on the basis of an expression sequence tag (EST) database search. We also found at least two additional candidates of this gene family in the database. These genes were induced by erythropoietin and granulocyte-macrophage colony stimulating factor in certain hematopoietic cell lines. The SH2 domain and a C-terminal 40 amino acid region, designated the CIS homology domain (CH domain), are highly conserved in this family, while the N-terminal regions of these proteins share little similarity. A yeast two-hybrid assay andin vitroandin vivobinding assays revealed that in addition to JAB, CIS3 bound to the JAK2 tyrosine kinase domain (JH1), although the interaction of CIS3 with the JAK2-JH1 domain was much weaker than that of JAB. Transient expression of JAB and CIS3, but not other CISs, strongly inhibited leukemia inhibitory factor (LIF)-induced STAT3-reporter gene activation in 293 cells. Furthermore, constitutive overexpression of JAB and CIS3 in M1 leukemia cells prevented LIF-induced differentiation and growth arrest. Although the physiological function remains to be investigated, CIS family genes could play a role in the negative regulation of cytokine signaling by interacting with specific targets.     

拟南芥转录因子GRAS家族基因群响应渗透和干旱胁迫的初步探索

GRAS家族是一类植物特有的转录调控因子,已有报道表明该家族基因在植物生长发育和光信号转导过程中具有重要作用.目前在拟南芥(Arabidopsis thaliana)基因组中已鉴定了33个GRAS家族基因.利用功能基因组学和生物信息学手段,通过基因芯片数据挖掘和基因功能预测,对拟南芥GRAS家族基因在渗透和干旱胁迫过程中的应答模式进行了初步探索,提出了一类响应渗透胁迫和干旱胁迫的拟南芥GRAS家族基因.以SCL13为例,利用基因芯片相关性和GO分析,对其在渗透胁迫信号转导过程中可能的调控机制进行了预测和分析.这一研究将为阐明GRAS家族基因参与水分胁迫的分子机制提供新的思路,同时也为植物抗逆分子育种提供候选基因.     

拟南芥转录因子GRAS 家族基因群响应渗透和干旱胁迫的初步探索

GRAS家族是一类植物特有的转录调控因子, 已有报道表明该家族基因在植物生长发育和光信号转导过程中具有重要作用。目前在拟南芥(Arabidopsis thaliana)基因组中已鉴定了33个GRAS家族基因。利用功能基因组学和生物信息学手段,通过基因芯片数据挖掘和基因功能预测, 对拟南芥GRAS家族基因在渗透和干旱胁迫过程中的应答模式进行了初步探索, 提出了一类响应渗透胁迫和干旱胁迫的拟南芥GRAS家族基因。以SCL13为例, 利用基因芯片相关性和GO分析, 对其在渗透胁迫信号转导过程中可能的调控机制进行了预测和分析。这一研究将为阐明GRAS家族基因参与水分胁迫的分子机制提供新的思路, 同时也为植物抗逆分子育种提供候选基因。     

酵母异源功能互补在植物基因克隆中的应用

酵母作为研究高等真核生物的一种重要的模式生物,不仅在生物信息学方面发挥着重要的作用,其丰富的突变体在其他生物基因克隆和功能验证等方面也发挥着重要的作用。酵母异源互补方法是利用酵母突变体验证相应性状异源基因功能,或通过文库筛选克隆相应性状异源基因,为高等真核生物的基因克隆和功能验证提供了一种捷径。主要对酵母异源功能互补法在研究植物基因方面的进展做一概述。     

Cloning and Functional Characterization of SAD Genes in Potato

Stearoyl-acyl carrier protein desaturase (SAD), locating in the plastid stroma, is an important fatty acid biosynthetic enzyme in higher plants. SAD catalyzes desaturation of stearoyl-ACP to oleyl-ACP and plays a key role in determining the homeostasis between saturated fatty acids and unsaturated fatty acids, which is an important player in cold acclimation in plants. Here, four new full-length cDNA of SADs (ScoSAD, SaSAD, ScaSAD and StSAD) were cloned from four Solanum species, Solanum commersonii, S. acaule, S. cardiophyllum and S. tuberosum, respectively. The ORF of the four SADs were 1182 bp in length, encoding 393 amino acids. A sequence alignment indicated 13 amino acids varied among the SADs of three wild species. Further analysis showed that the freezing tolerance and cold acclimation capacity of S. commersonii are similar to S. acaule and their SAD amino acid sequences were identical but differed from that of S. cardiophyllum, which is sensitive to freezing. Furthermore, the sequence alignments between StSAD and ScoSAD indicated that only 7 different amino acids at residues were found in SAD of S. tuberosum (Zhongshu8) against the protein sequence of ScoSAD. A phylogenetic analysis showed the three wild potato species had the closest genetic relationship with the SAD of S. lycopersicum and Nicotiana tomentosiformis but not S. tuberosum. The SAD gene from S. commersonii (ScoSAD) was cloned into multiple sites of the pBI121 plant binary vector and transformed into the cultivated potato variety Zhongshu 8. A freeze tolerance analysis showed overexpression of the ScoSAD gene in transgenic plants significantly enhanced freeze tolerance in cv. Zhongshu 8 and increased their linoleic acid content, suggesting that linoleic acid likely plays a key role in improving freeze tolerance in potato plants. This study provided some new insights into how SAD regulates in the freezing tolerance and cold acclimation in potato.     

Transformation of Two Nitrogen-fixation-related Plant Genes into Tobacco and Their Expressions

Lectins and leghemoglobins in legumes play the important roles, respectively, in recognition of host plants to their own rhizobia, and lowering the oxygen partial pressure surround the bacteroids and protecting nitrogenase from oxygen in symbiotic nitrogen-fixing nodules.In order to investigate the non-leguminous recognition of rhizobial bacteria relating to nitrogen fixation, plant expression vectors containing pea lectin gene (pl) and Parasponia hemoglobin gene (phb) have been, respectively, constructed in a plasmid and the plasmid has been introduced into tobacco (Nicotiana tabacum L.) using Agrobacterium tumefaciens (Smith et Townsend) Conn as a vehicle for transformation. PCR and Southern blot demonstrated that the two genes were integrated into the genome of the tobacco plants. Histochemical staining for GUS activity, Western blotting,and in situ hybridization of pea lectin showed that they were expressed at translational level in the plants. These results may provide a clue for exploring whether Rhizobium leguminosarum bv. viciae could extend its host range and make the transgenic tobacco plants have the possibility of being symbiotic, or associative to nitrogen fixation.     

水稻穗发育功能基因对穗期氮肥的响应

为探讨氮肥对水稻(Oryza sativa)穗发育的调控作用, 使用高通量测序技术检测氮肥处理前后水稻叶片和幼穗组织中转录组的变化, 并从中筛选到大量差异表达基因。这些基因的功能涉及转录调控、激素代谢和信号转导、物质代谢和转运、胁迫响应、信号转导(受体)和蛋白质降解等。同时对目前克隆得到的穗发育相关基因进行分析, 发现在氮素穗肥的作用下, 部分重要功能基因的表达量发生了明显变化, 其中一些基因还参与调控水稻株高、抽穗期、分蘖和结实率等性状。对这些差异表达基因的功能研究有助于揭示氮素穗肥调控水稻每穗颖花数的分子机制。