In planta characterization of a tau class glutathione S-transferase gene from <Emphasis Type="Italic">Juglans regia</Emphasis> (<Emphasis Type="Italic">JrGSTTau1</Emphasis>) involved in chilling tolerance

Key message

JrGSTTau1 is an important candidate gene for plant chilling tolerance regulation.

Abstract

A tau subfamily glutathione S-transferase (GST) gene from Juglans regia (JrGSTTau1, GeneBank No.: KT351091) was cloned and functionally characterized. JrGSTTau1 was induced by 16, 12, 10, 8, and 6 °C stresses. The transiently transformed J. regia showed much greater GST, glutathione peroxidase (GPX), superoxide dismutase (SOD), and peroxidase (POD) activities and lower H₂O₂, malondialdehyde (MDA), reactive oxygen species (ROS), and electrolyte leakage (EL) rate than prokII (empty vector control) and RNAi::JrGSTTau1 under cold stress, indicating that JrGSTTau1 may be involved in chilling tolerance. To further confirm the role of JrGSTTau1, JrGSTTau1 was heterologously expressed in tobacco, transgenic Line5, Line9, and Line12 were chosen for analysis. The germinations of WT, Line5, Line9, and Line12 were similar, but the fresh weight, primary root length, and total chlorophyll content (tcc) of the transgenic lines were significantly higher than those of WT under cold stress. When cultivated in soil, the GST and SOD activities of transgenic tobacco were significantly higher than those of WT; however, the MDA and H₂O₂ contents of WT were on average 1.47- and 1.96-fold higher than those of Line5, Line9, and Line12 under 16 °C. The DAB, Evans blue, and PI staining further confirmed these results. Furthermore, the abundances of NtGST, MnSOD, NtMAPK9, and CDPK15 were elevated in 35S::JrGSTTau1 tobacco compared with WT. These results suggested that JrGSTTau1 improves the plant chilling tolerance involved in protecting enzymes, ROS scavenging, and stress-related genes, indicating that JrGSTTau1 is a candidate gene for the potential application in molecular breeding to enhance plant abiotic stress tolerance.

     

Systematic revision of mountain dragons (Reptilia: Agamidae: Diploderma) in China,with descriptions of six new species and discussion on their conservation

Cryptic diversity not only introduces confusion to taxonomic studies, but it also poses major challenges to conservation and environmental legislation. One such troubling group are the Mountain Dragons in the genus Diploderma in Southwest China. Previous studies have suggested that the genus contains considerable cryptic diversity, particularly in the D. flaviceps complex. Owing to taxonomic confusion, micro‐endemic lineages are still neglected by the Chinese wildlife protection laws, despite their urgent conservation needs. Combining multivariate morphological and multi‐locus phylogenetic data, we provide the first integrative systematic revision of the genus Diploderma. Specifically, we confirm that the six examined populations of D. cf. flaviceps from the upper Jinsha and Yalong River Valleys in Sichuan and Yunnan Provinces represent six cryptic, undescribed lineages, and we describe each of them as a new species. With the updated taxonomy and distribution information, we discuss the taxonomy of the D. flaviceps complex in Southwest China, provide an updated diagnostic key along with distributional ranges for all species of the genus, and discuss some of the suspicious records of other congeners in China. Lastly, we evaluate the IUCN status of each of the six new species and highlight the major challenges for Diploderma conservation in China due to delayed environmental legislation and misleading conservation assessments.     

Patterning parameters associated with the branching of the ureteric bud regulated by epithelial-mesenchymal interactions

The mechanisms by which the branching of epithelial tissue occurs and is regulated to generate different organ structures are not well understood. In this work, image analyses of the organ rudiments demonstrate specific epithelial branching patterns for the early lung and kidney; the lung type typically generating several side branches, whereas kidney branching was mainly dichotomous. Parameters such as the number of epithelial tips, the angle of the first branch, the position index of the first branch (PIFB) in a module, and the percentage of epithelial module type (PMT) were analysed. The branching patterns in the cultured lung and kidney, and in homotypic tissue recombinants recapitulated their early in vivo branching patterns. The parameters were applied to heterotypic tissue recombinants between lung mesenchyme and ureteric bud, and tip number, PIFB and PMT values qualified the change in ureter morphogenesis and the reprogramming of the ureteric bud with lung mesenchyme. All the values for the heterotypic recombinant between ureteric bud and lung mesenchyme were significantly different from those for kidney samples but similar to those of the lung samples. Hence, lung mesenchyme can instruct the ureteric bud to undergo aspects of early lung-type epithelial morphogenesis. Different areas of the lung mesenchyme, except the tracheal region, were sufficient to promote ureteric bud growth and branching. In conclusion, our findings provide morphogenetic parameters for monitoring epithelial development in early embryonic lung and kidney and demonstrate the use of heterotypic tissue recombinants as a model for studying tissue-specific epithelial branching during organogenesis.     

安徽天柱山夏末开花植物的花粉形态

通过光镜和扫描电镜对天柱山夏末开花的23科、39属、37种3变种的植物花粉形态进行了观察研究.从形状看,一些科内花粉形状具有一致性,如菊科[Asteraceae Dumortier (Compositae Giseke,nom.altern.)]6个种都是球形或近球形(P/E=0.95-1.03);豆科(Legumin...     

Mesophyll conductance variability of rice aquaporin knockout lines at different growth stages and growing environments

The plasma membrane subfamily of aquaporins [plasma membrane intrinsic proteins (PIPs)], which facilitates the CO₂ diffusion across membranes, is proposed to play an important role in mesophyll conductance to CO₂ (g_m), a major limiting factor of photosynthesis. However, recent studies implied no causal relationship between g_m and PIPs because they failed to repeat the previous observed differences in g_m between PIP knockout lines and their wild-type. The contrasting results on the role of PIPs in g_m were interpreted as the different growth conditions among studies, which has never been tested. Here, we assessed whether the differences in g_m between wild-type and PIP knockout lines, Ospip1;1, Ospip1;2 and Ospip2;1, varied with growth condition (field versus pot condition) and growth stages in rice. Under field conditions, no differences were observed in plant performance, photosynthetic rate (A) and g_m between PIP knockout lines and the wild-type. However, in agreement with previous studies, two out of three knockout lines showed significant declines in tiller number, plant height, A and g_m under pot conditions. Moreover, we found that the differences in A and g_m between PIP knockout lines and the wild-type varied with the growth stage of the plants. Our results showed that the differences in g_m between PIP knockout lines and wild-type were depending on the growth environments and stage of the plants, and further efforts are required to reveal the underlying mechanisms.     

Over-expression of <Emphasis Type="Italic">JrsHSP17.3</Emphasis> gene from <Emphasis Type="Italic">Juglans regia</Emphasis> confer the tolerance to abnormal temperature and NaCl stresses

Small heat shock proteins (sHSPs) are an HSP subgroup and involved in environmental stress response. In the current study, to understand the role of sHSP protein in a widely distribution nut woody tree, a sHSP gene was cloned from Juglans regia (JrsHSP17.3, GeneBank No.: KT277704). Compared with control condition, the expression of JrsHSP17.3 was induced to 58.1-fold (6 h) in the roots, 86.8-fold in the stems (9 h), 50.9-fold in the leaves (6 h) under 10°C; and was up-regulated to 2.9- ～ 79.9-fold response to 40°C for 3～9 h; meanwhile, it was transcribed to 5.9 - ～39.7-fold under 9 h NaCl treatment, suggesting the potential role of JrsHSP17.3 to cold, heat and NaCl stimulus. Further, JrsHSP17.3 transgenic yeasts showed improved tolerance to freezing, heat and salt stresses compared with control yeast. JrsHSP17.3 was transient over-expressed in J. regia leaves. The leaves non-transgenic (NT) and vector prokII transgenic (empty, PT) were used as control. The expression of JrsHSP17.3 was 81.6-, 125.4-, and 54.2-fold of the control lines under normal conditions, indicating the success over-expression of JrsHSP17.3. Cell damage staining and physiological index determination showed that JrsHSP17.3 transformed lines, NT and PT displayed no obvious difference under control conditions, however, after treated with 16°C, 40°C and NaCl, JrsHSP17.3 transformed lines displayed weaker cell damage, lower level of electrolyte leakages (EL) rate, malondialdehyde (MDA) and H₂O₂ content, and higher activities of catalase (CAT), glutathione transferase (GST), superoxide dismutase (SOD) and peroxidase (POD) as well as more accumulation of proline than NT and PT. Meanwhile, NT and PT were similar and showed no significant difference under all conditions. All of these results indicated that JrsHSP17.3 can improve plant tolerance to abnormal temperatures and NaCl stresses, it represents a potential candidate gene for molecular breeding to enhance stress tolerance in plants.     

RNA干扰技术及其在植物研究中的应用

RNA干扰(RNA interference,RNAi)是最近几年发现和发展起来的一门新兴的在转录水平上的基因阻断技术,它是生物体内由双链RNA(double-stranded RNA,dsRNA)介导同源mRNA降解的现象。RNAi广泛存在于从真菌到高等植物、从无脊椎动物到哺乳动物各种生物中。研究表明通过转入目的基因序列的双链RNA可以诱导产生基因沉默现象。同时,RNAi能监控异常的或外源的遗传物质在机体内的水平,并调控基因的表达,是生物体抵御外在感染的一种重要的保护机制,这使得RNA干扰技术具有十分诱人的应用前景。介绍了RNAi的研究历史、作用机制、特点及其在植物研究中的应用。