Chenghao Li

Tamarix hispida metallothionein-like ThMT3, a reactive oxygen species scavenger, increases tolerance against Cd(2+), Zn(2+), Cu(2+), and NaCl in transgenic yeast.

A metallothionein-like gene, ThMT3, encoding a type 3 metallothionein, was isolated from a Tamarix hispida leaf cDNA library. Expression analysis revealed that mRNA of ThMT3 was upregulated by high salinity as well as by heavy metal ions, and that ThMT3 was predominantly expressed in the leaf. Transgenic yeast (Saccharomyces cerevisiae) expressing ThMT3 showed increased tolerance to Cd2+, Zn2+, Cu2+, and NaCl stress. Transgenic yeast also accumulated more Cd2+, Zn2+, and NaCl, but not Cu2+. Analysis of the expression of four genes (GLR1, GTT2, GSH1, and YCF1) that aid in transporting heavy metal (Cd2+) from the cytoplasm to the vacuole demonstrated that none of these genes were induced under Cd2+, Zn2+, Cu2+, and NaCl stress in ThMT3-transgenic yeast. H2O2 levels in transgenic yeast under such stress conditions were less than half those in control yeast under the same conditions. Three antioxidant genes (SOD1, CAT1, and GPX1) were specifically expressed under Cd2+, Zn2+, Cu2+, and NaCl stress in the transgenic yeast. Cd2+, Zn2+, and Cu2+ increased the expression levels of SOD1, CAT1, and GPX1, respectively, whereas NaCl induced the expression of SOD1 and GPX1.

Genome-Wide Analysis of C2H2 Zinc-Finger Family Transcription Factors and Their Responses to Abiotic Stresses in Poplar (Populus trichocarpa).

Background C2H2 zinc-finger (C2H2-ZF) proteins are a large gene family in plants that participate in various aspects of normal plant growth and development, as well as in biotic and abiotic stress responses. To date, no overall analysis incorporating evolutionary history and expression profiling of the C2H2-ZF gene family in model tree species poplar (Populus trichocarpa) has been reported. Principal Findings Here, we identified 109 full-length C2H2-ZF genes in P. trichocarpa, and classified them into four groups, based on phylogenetic analysis. The 109 C2H2-ZF genes were distributed unequally on 19 P. trichocarpa linkage groups (LGs), with 39 segmental duplication events, indicating that segmental duplication has been important in the expansion of the C2H2-ZF gene family. Promoter cis-element analysis indicated that most of the C2H2-ZF genes contain phytohormone or abiotic stress-related cis-elements. The expression patterns of C2H2-ZF genes, based on heatmap analysis, suggested that C2H2-ZF genes are involved in tissue and organ development, especially root and floral development. Expression analysis based on quantitative real-time reverse transcription polymerase chain reaction indicated that C2H2-ZF genes are significantly involved in drought, heat and salt response, possibly via different mechanisms. Conclusions This study provides a thorough overview of the P. trichocarpa C2H2-ZF gene family and presents a new perspective on the evolution of this gene family. In particular, some C2H2-ZF genes may be involved in environmental stress tolerance regulation. PtrZFP2, 19 and 95 showed high expression levels in leaves and/or roots under environmental stresses. Additionally, this study provided a solid foundation for studying the biological roles of C2H2-ZF genes in Populus growth and development. These results form the basis for further investigation of the roles of these candidate genes and for future genetic engineering and gene functional studies in Populus.

Genome-wide identification and expression profiling of the copper transporter gene family in Populus trichocarpa.

Copper transporters (COPT/Ctr) have important roles in the transport of copper (Cu) across the cell membrane in many different species. A comprehensive phylogeny and a molecular structure analysis of the COPT/Ctr family in plants and animals are presented, with an emphasis and bioinformatic analysis of the copper transporter family in Populus trichocarpa (PtCOPT). Structural analyses of PtCOPTs showed that most have 3 transmembrane domains (TMDs), with an exception of PtCOPT4 (2 TMDs). Gene structure, gene chromosomal location, and synteny analyses of PtCOPTs demonstrated that tandem and segmental duplications have likely contributed to the expansion and evolution of the PtCOPTs. Additionally, promoter analyses showed that the function of PtCOPTs is related to Cu and ferrum (Fe) transport. Tissue-specific expression of PtCOPT genes showed that most had relatively high transcript levels in roots and leaves. Quantitative real-time RT-PCR (qRT-PCR) analysis revealed that the expression of PtCOPT genes were induced not only in limited and excessive Cu, Fe, zinc (Zn) and manganese (Mn) stress, but also in lead (Pb), and cadmium (Cd) stress.

Overexpression of TaLEA3 induces rapid stomatal closure under drought stress in Phellodendron amurense Rupr

Late embryogenesis abundant (LEA) proteins participate in drought stress responses in plants. In the present study, the gene TaLEA3 from the drought-resistant plant Tamarix androssowii was transformed into Amur cork tree (Phellodendron amurense) via Agrobacterium tumefaciens to investigate the mechanism of stomatal closure in response to osmotic stress. Our results showed that P. amurense overexpressing TaLEA3 were resistant to drought stress by rapid stomatal closure. To study the stomatal movement regulated at the molecular level, a model system for stoma closure was established in in vitro P. amurense. In this work, we found that the increased Ca2+ accumulation in guard cells of transgenic plants caused stomatal closure and activated K+ efflux under polyethylene glycol (PEG) stress. Moreover, H+ changes might provide a needed pH condition for stomatal closure. Further, nitric oxide (NO) fluorescence was measured using an NO-specific fluorescent probe, diaminofluorescein-FM diacetate, which showed that guard cell NO fluorescence was stronger in transgenic plants compared with wild type plants. Additionally, five genes encoding nitrate reductase were up-regulated, indicating that TaLEA3 overexpression positively regulated NO biosynthesis and accumulation in the guard cells. This discovery will further our understanding of the LEA gene function and will help in engineering drought-resistant tree cultivars.

Genome-Wide Analysis of the ZRT, IRT-Like Protein (ZIP) Family and Their Responses to Metal Stress in Populus trichocarpa

The ZRT, IRT-like protein (ZIP) family plays an important role in the transport of zinc (Zn) and iron (Fe) across the cell membrane in many different species. However, studies on ZIP family are mainly limited in herbaceous species; hence, we investigated functional divergence of ZIP family in Populus trichocarpa. We identified 21 ZIP genes in P. trichocarpa and classified them into four groups based on phylogenetic analysis. Structural analyses revealed that most of the PtrZIP transporters have eight transmembrane domains (TMDs). PtrZIP members were unequally positioned in 19 P. trichocarpa linkage groups (LGs), with six tandem duplications and four segmental duplications. The promoter regions of PtrZIP genes contain Zn, Fe, copper (Cu), and other metal stress-related cis-elements. Additionally, tissue-specific expression of PtrZIP genes showed that most of them had relatively high expression levels in the root. Quantitative real-time RT-PCR (qRT-PCR) analysis revealed that the expression of PtrZIP genes were induced not only under deficiency or excess condition of Zn, Fe, Cu and manganese (Mn) but also under excess condition of cadmium (Cd) and lead (Pb) stress. These findings indicated that PtrZIP genes may have played potential roles in metal transporters. Genome-wide analysis of PtrZIP genes in P. trichocarpa provided more comprehensive insights on the structure and function of this gene family.