13C脉冲标记法：不同生育期水稻光合碳在植物-土壤系统中的分配

定量生育期内植物光合碳在植物组织-土壤的分配规律,对于理解全球碳循环有着重要意义。采用~(13)C-CO_2脉冲标记结合室内培养,通过元素分析仪-稳定同位素联用(Flash HT-IRMS)分析植物各部分及土壤δ~(13)C值,比较了不同生育期下水稻光合碳在不同组织间的分配规律,并量化了水稻光合碳向土壤碳库的转移。结果表明:(1)水稻地上部和根系干物质量随水稻生育期的增加而呈现递增趋势,不同的生育期表现为:分蘖期拔节期抽穗期扬花期成熟期。而整个生育期的根冠比为0.2—0.4,分蘖期的根冠比最高,随着水稻生育期的增加而递减,到抽穗期以后根冠比稳定在0.2左右。(2)脉冲标记6h后,水稻地上部和地下部(根系)的δ~(13)C值在-25.52‰—-28.33‰,不同器官的δ~(13)C值存在明显分馏效应,且趋势基本一致,即茎杆(籽粒)叶片(根系);这种由于水稻生育期特性导致的各器官碳同位素分馏的现象,可用于指示不同生育期下水稻光合碳的分配和去向。(3)不同生育期~(13)C-光合碳在植物-土壤系统的分配规律不同,生长前期光合碳向根系及土壤中分配的比例高,具有较强的碳汇能力,而随生育期光合碳在根系及土壤中的分配比例呈下降趋势,但积累量不断增加。(4)不同生育期~(13)C光合碳在水稻-土壤系统中的分配比例差异明显。水稻分蘖期有近30%光合碳用于根系建成并部分通过根系分泌物进入土壤有机碳库(10%),而到成熟期则向籽粒中分配较多,而且光合碳在土壤中的分配比例也随生育期呈下降趋势。研究结果对稻田土壤有机碳循环过程和调控机制的揭示具有重要的理论意义。     

Relative importance of osmotic-stress and ion-specific effects on ABA-mediated inhibition of leaf expansion growth in Phaseolus vulgaris

The osmotic and ion-specific components of salt-induced inhibition of leaf expansion growth were investigated in beans grown from 12 h to several days in either NaCl-containing solution cultures, an isosmotic concentrated macronutrient solution, or a vermiculite–compost mixture with low Na⁺ but high Cl^– availability. Inhibition of leaf expansion and leaf ABA increase was more intense in the NaCl than in the isosmotic macronutrient treatment. Root Na⁺ was highly correlated to inhibition of leaf expansion and leaf or xylem sap ABA. When Na⁺ was sequestered in soil, salinized plants showed no reduction in leaf expansion or ABA increase, regardless of the presence of high leaf Cl^– concentrations. Stomatal conductance exhibited an exponential relationship with the reciprocal value of xylem sap ABA. Our results indicate that an ion-specific effect caused by Na⁺ in roots may account for an ABA-mediated reponse of both stomatal closure and leaf expansion inhibition.     

Deoxyribonuclease 1-like 3 Inhibits Hepatocellular Carcinoma Progression by Inducing Apoptosis and Reprogramming Glucose Metabolism

HCC has remained one of the challenging cancers to treat, owing to the paucity of drugs targeting the critical survival pathways. Considering the cancer cells are deficient in DNase activity, the increase of an autonomous apoptisis endonuclease should be a reasonable choice for cancer treatment. In this study, we investigated whether DNASE1L3, an endonuclease implicated in apoptosis, could inhibit the progress of HCC. We found DNASE1L3 was down-regulated in HCC tissues, whereas its high expression was positively associated with the favorable prognosis of patients with HCC. Besides, serum DNASE1L3 levels were lower in HCC patients than in healthy individuals. Functionally, we found that DNASE1L3 inhibited the proliferation of tumor cells by inducing G0/G1 cell cycle arrest and cell apoptosis in vitro. Additionally, DNASE1L3 overexpression suppressed tumor growth in vivo. Furthermore, we found that DNASE1L3 overexpression weakened glycolysis in HCC cells and tissues via inactivating the rate-limiting enzymes involved in PTPN2-HK2 and CEBPβ-p53-PFK1 pathways. Finally, we identified the HBx to inhibit DNASE1L3 expression by up-regulating the expression of ZNF384. Collectively, our findings demonstrated that DNASE1L3 could inhibit the HCC progression through inducing cell apoptosis and weakening glycolysis. We believe DNASE1L3 could be considered as a promising prognostic biomarker and therapeutic target for HCC.     

Agrobacterium-mediated transformation of Asparagus officinalis L.: molecular and genetic analysis of transgenic plants

Four long-term embryogenic lines of Asparagus officinalis were co-cultured with the hypervirulent Agrobacterium tumefaciens strain AGL1Gin carrying a uidA gene and an nptII gene. 233 embryogenic lines showing kanamycin resistance and -glucuronidase (GUS) activity were obtained. Transformation frequencies ranged from 0.8 to 12.8 transformants per gram of inoculated somatic embryos, depending on the line. Southern analysis showed that usually 1 to 4 T-DNA copies were integrated. Regenerated plants generally exhibited the same insertion pattern as the corresponding transformed embryogenic line. T₁ progeny were obtained from crosses between 6 transformed plants containing 3 or 4 T-DNA copies and untransformed plants. They were analysed for GUS activity and kanamycin resistance. In three progenies, Mendelian 1:1 segregations were observed, corresponding to one functional locus in the parent transgenic plants. Southern analysis confirmed that T-DNA copies were inserted at the same locus. Non-Mendelian segregations were observed in the other three progenies. T₂ progeny also exhibited non-Mendelian segregations. Southern analysis showed that GUS-negative and kanamycin-sensitive plants did not contain any T-DNA, and therefore inactivation of transgene expression could not be responsible for the abnormal segregations.     

Maximizing antibody production in a targeted integration host by optimization of subunit gene dosage and position

Historically, therapeutic protein production in Chinese hamster ovary (CHO) cells has been accomplished by random integration (RI) of expression plasmids into the host cell genome. More recently, the development of targeted integration (TI) host cells has allowed for recombination of plasmid DNA into a predetermined genomic locus, eliminating one contributor to clone-to-clone variability. In this study, a TI host capable of simultaneously integrating two plasmids at the same genomic site was used to assess the effect of antibody heavy chain and light chain gene dosage on antibody productivity. Our results showed that increasing antibody gene copy number can increase specific productivity, but with diminishing returns as more antibody genes are added to the same TI locus. Random integration of additional antibody DNA copies in to a targeted integration cell line showed a further increase in specific productivity, suggesting that targeting additional genomic sites for gene integration may be beneficial. Additionally, the position of antibody genes in the two plasmids was observed to have a strong effect on antibody expression level. These findings shed light on vector design to maximize production of conventional antibodies or tune expression for proper assembly of complex or bispecific antibodies in a TI system.     

Hydrotropism and its interaction with gravitropism in roots

We have studied hydrotropism and its interaction with gravitropism in agravitropic roots of a pea mutant and normal roots of peas (Pisum sativum L.) and maize (Zea mays L.). The interaction between hydrotropism and gravitropism in normal roots of peas or maize were also examined by nullifying the gravitropic response on a clinostat and by changing the stimulus-angle for gravistimulation. Depending on the intensity of both hydrostimulation and gravistimulation, hydrotropism and gravitropism of seedling roots strongly interact with one another. When the gravitropic response was reduced, either genetically or physiologically, the hydrotropic response of roots became more unequivocal. Also, roots more sensitive to gravity appear to require a greater moisture gradient for the induction of hydrotropism. Positive hydrotropism of roots occurred due to a differential growth in the elongation zone; the elongation was much more inhibited on the moistened side than on the dry side of the roots. It was suggested that the site of sensory perception for hydrotropism resides in the root cap, as does the sensory site for gravitropism. Furthermore, an auxin inhibitor, 2,3,5-triiodobenzoic acid (TIBA), and a calcium chelator, ethyleneglycol-bis-(-aminoethylether)-N,N,N,N- tetraacetic acid (EGTA), inhibited both hydrotropism and gravitropism in roots. These results suggest that the two tropisms share a common mechanism in the signal transduction step.     

Evidence for fine-scale habitat specialisation in an invasive weed

Aims As an exotic species colonises a new continent, it must overcome enormous environmental variation in its introduced range. Local adaptation of introduced species has frequently been observed at the continent scale, particularly in response to latitudinal climatic variation. However, significant environmental heterogeneity can also exist at the landscape scale. A small number of studies have provided evidence that introduced species may also be capable of phenotypic and genetic differentiation at much smaller spatial scales. For example, previously we found US agricultural and non-agricultural populations of Sorghum halepense (Johnsongrass) to be phenotypically and genetically distinct. In this study, we investigated whether this phenotypic differentiation of agricultural and non-agricultural populations of S. halepense is the result of fine-scale local specialisation.Methods We surveyed a nationally collected S. halepense germplasm panel and also collected neighbouring agricultural and non-agricultural sub-populations of S. halepense at four sites throughout Western Virginia, USA, raising seedlings in common conditions mimicking both agricultural and non-agricultural habitats.Important findings At the national scale, we found evidence of habitat differentiation but not specialisation. However, at the local scale, we found evidence of specialisation in two of the four local populations to non-agricultural habitat, but no evidence of specialisation to agricultural habitat. These results show that local specialisation is a possible, but not guaranteed consequence of kilometre-scale habitat heterogeneity in invasive species. This finding contributes to a growing awareness of the importance of fine-scale local adaptation in the ecology and management of introduced and weedy species.     

Tolerance ofFestuca rubra L. to zinc in relation to mycorrhizal infection

Summary Plant yield of mycorrhizal and non-mycorrhizalFestuca rubra L. was linearly decreased with increasing zinc concentrations in nutrient solution. In all cases, non-mycorrhizal plant growth was significantly greater than that of mycorrhizal plants. Zinc and phosphorus concentrations of root and shoot of mycorrhizal plants were greater in all zinc treatments while mycorrhizal plants showed equal or lower tolerance indices to zinc than non-mycorrhizal plants. Yield depressions of mycorrhizal plants may be the result of enhanced zinc and phosphorus concentrations combined with the cost for growth and maintenance of the mycorrhizal fungi.