Cloning of 9-cis-epoxycarotenoid dioxygenase （NCED） gene encoding a key enzyme during abscisic acid （ABA） biosynthesis and ABA-regulated ethylene production in detached young persimmon calyx

Unlike the typical climacteric fruits, persimmons （Diospyros kaki Thunb.） produce higher levels of ethylene when they are detached from trees at a younger stage. In order to obtain detailed information on the role of abscisic acid （ABA） in ripening, we cloned the DKNCED1, DKACS2, and DKACO1 genes from the calyx. Water loss was first noted in the calyx lobe, and DKNCED1 was highly expressed 1 d after the fruits were detached, coinciding with an increase in the ABA content. Then, the DKACS2 and DKACO1 genes were expressed after some delay. In the calyx, the ABA peak was observed 2 d after the fruits were harvested, and this peak preceded the ethylene peak observed on day 3. The fruit firmness rapidly decreased on day 4, and the fruits softened completely 6 d after they were harvested. The increases in the expressions of ABA, ethylene, and the genes in the calyxes occurred earlier than the corresponding increases in the pulp, although the 3 increases occurred on different days. Exogenous ABA treatment increased ABA concentration, induced expression of both ACS and ACO, and promoted ethylene synthesis and young-fruit softening; by contrast, treatment with NDGA inhibited the gene expressions and ethylene synthesis and delayed young-fruit softening. These results indicate that ethylene biosynthesis in the detached young persimmon fruits is initially triggered by ABA, which is induced by water loss in the calyx, through the induction of DKACS2 and DKACO1 expressions. The ethylene produced in the calyx subsequently diffuses into the pulp tissue, where it induces autocatalytic ethylene biosynthesis, resulting in an abrupt increase in ethylene production.     

Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

The mechanisms for the production of hydrogen peroxide （H2O2） induced by abscisic acid （ABA） were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H2O2, which was mainly derived from super-oxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane （PM） NADPH oxidase activity, which coin- cided on time and magnitude with the elevation in ABA-induced accumulation of H2O2. Moreover, these enhanced effects were pro- nouncedly inhibited by two NADPH oxidase inhibitors, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of H2O2 in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H2O2 in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in sus-pension culture cells of tobacco, and that NADPH oxidase and H2O2 appear to be important components in ABA signal transduction pathway in plants.     

Development and formability analysis of TRIP seamless steel tube

In this paper, the production technology of transformation induced plasticity （TRIP） steel was first introduced into the steel tube manufacture field to produce the steel tubes with high strength and plasticity. The TRIP seamless steel tubes with the microstructure of ferrite, bainite, retained austenite and a little martensite were successfully fabricated using a cold-drawn steel tube with two-stage heat treatment technique and continu- ous heat treatment process, respectively. The ring tensile test and cold bend test were carried out to study the formability of the newly developed TRIP seamless steel tube. The results showed that the TRIP seamless steel tubes have a good cold formability, and they are available to be used in the tube hydroforming process. In ad- dition, the equipment of continuous heat treatment developed in the current study can be used to produce TRIP steel tube, and it may serve as an important reference for the industrial production of TRIP steel tube.     

FAILURE ANALYSIS OF THE COLD FORGING DIE USED FOR COPPER CONTACT-HEAD MANUFACTURE

The lifetime of dies is an important factor in raising the economic efficiency in production.Through the microscopic analysis as well as metalloscopy on the failure status of the cold forgingdie destined,for manufacture of copper contact heads of electric switches,the paper reaches theconclusion that the inhomogeneous distribution of carbides will cause fatigue breaking.And themeasures to control the quality of material and to improve the heat treatment technology are pro-posed.     

The entransy dissipation minimization principle under given heat duty and heat transfer area conditions

Under given heat duty and heat transfer area conditions, the equipartition of the entransy dissipation (EoED) principle, the equipartition of the temperature difference (EoTD) principle, and the equipartition of the heat flux (EoHF) principle are applied to the optimization design of a heat exchanger with a variable heat transfer coefficient. The results show that the difference between the results obtained using the EoED and EoTD principles is very small, far smaller than that between the results obtained using the EoED and EoHF principles. The correct entransy dissipation minimization principle is chosen to optimize the parameters in the hot and cold fluids in a two-fluid heat exchanger, under given heat duty and heat transfer area conditions. The results indicate that the proper choice of the two alternative fluids has an important role in the successful application of the entransy dissipation minimization principle. The fluid that could improve the total heat transfer coefficient should be chosen, or the fluid that makes the temperature profiles of the hot and cold fluids parallel and decreases the temperature difference between the hot and cold fluids after optimization simultaneously, could be the proper one.     

Cyclic AMP response element binding protein (CREB) participates in the heat-inducible expression of humanhsp90β gene

Human heat shock protein 90b gene ( hsp90b ) is a constitutively expressed heat shock gene existing in most of cell types tested that can be further induced by heat shock. Chloramphenical acetyl transferase (CAT) reporter plasmids driven by different regulatory fragments of hsp90b gene were constructed and transfected into Jurkat cells to explore the role of a cAMP response element (CRE) in the upstream of the gene. Results show that, in comparison with the wild type construct, a severe reduction (~2/3) in the increased folds of promoter activity induced by heat shock at 42℃ for 1 h was observed in a construct with CRE-containing fragment (-173/-91bp) deleted. Electrophoretic mobility shift assays (EMSA) showed that phosphorylated CRE-binding protein (CREB) in the nuclear extract of heat shocked Jurkat cells is specifically bound to the fragment. Additionally, both of the phosphorylation on CREB and the activity of protein kinase A (PKA) were found in Jurkat cells to be enhanced with extending time of heat shock treatment. Our results indicate that in addition to the intronic HSE/HSF pathway, phosphorylated CREB also participates in the heat shock induced expression of human hsp90b gene via its interaction with CRE which may be regulated by PKA-sig- naling pathway.     

Measurement of ethylene and methane production in a temperate forest soil using inhibition of acetylene and carbon monoxide

We studied in the laboratory the effects of acetylene （C2H2） concentrations on the accumulation and consumption of ethylene and methane in a temperate pine forest soil, and in situ ethylene and methane production and flush effects of nitrogen sources on both productions in the pine forest stand （Pinus sylvestris L.）. The addition of C2H2 at concentrations more than 50 Pa C2H2 in the headspace caused a more than 95% reduction in rates of ethylene and methane consumption in forest soil compared to those with no C2H2. Furthermore, addition of acetylene within a range of 50 to 10, 000 Pa C2H2 induced a similar rate of methane accumulation in forest soil. Hence, it can be concluded that presence of more than 50 Pa C2H2 in the headspace is an effective method to measure methane production in forest soil. The addition of C2H2 at concentrations more than 50 Pa C2H2 induced an increasing concentration of ethylene in the headspace （P≤0.05）, indicating the reduction of acetylene to ethylene in forest soil. Using inhibition of 0.5 kPa C2H2 in combination with 5 kPa carbon monoxide that inhibits the reduction of acetylene in a short term, it was observed that there was a larger in situ methane production rate （218±26 μg C m^-2 h^-1（μg C per square meter per hour, the same below）） than in situ ethylene production rate （92±6 μg C m^-2 h^-1） in the pine forest soil. The addition of nitrogen sources such as urea, urea plus a nitrification inhibitor dicyandiamide, and potassium nitrate, could induce a 5-fold greater increase in rates of in situ ethylene and methane production compared to those in the control, particularly in the latter （P≤0.05）. The results can promote in situ measurement of ethylene and methane production in forest soils at different sites.     

Mathematical Model of ComputerHeat Treatment and Its Simulation

Computer simulation on heat treatment is the foundation of intelligent heat treatment. The simulations of temperature field,phase transformation, stress/strain complicate quenching operation were realized by using the model of three dimensional non-linear finite element method and the treatment methods of abruptly changing interface conditions. The simulation results basically fit those measured in experiments. The intelligent sealed multipurpose furnace production line has been developed based on the combination of computer simulation on gaseous carburizing and computer control technology. More than 3000 batches of workpieces have been processed on this production line, and all are up to standard. The application of computer simulation technology can significantly improve the loading ability and reliability of nitriding and carburizing workpieces, reduce heat treatment distortion, and shorten carburizing duration. It is recommended that the reliable product design without redundancy should be performed with the combination of the CAD of mechanical products, the CAE of materials selection and heat treatment, and the dynamic evaluation technology of product reliability.     

Characterization of a PDR type ABC transporter gene from wheat （Triticum aestivum L.）

DON, as a virulence factor, plays an important role in the infection of Fusarium graminearum in wheat. The infection ability of F. graminearum depends on its capacity of producing DON. The production of DON by F. graminearum is significantly decreased in the wheat varieties with scab resistance. In this study, GeneChip analysis indicated that an EST encoding an ATP-binding cassette （ABC） transporter was up-regulated by 45 times in a wheat landrace Wangshuibai, which is resistant to DON accumulation. A pair of EST-derived primers were designed based on the EST sequence, and a clone was then isolated from a wheat genomic DNA TAC library. The TAC clone was sequenced using chromosome walking and gene prediction was conducted using Softberry. A cDNA clone of this gene was subsequently isolated from Wangshuibai induced by DON using gene-specific primers designed according to the untranslated sequence of the gene. The genome size of the gene is 7377 bp, consisting of 19 exons with coding sequences of 4308 bp. It encodes a protein with 1435 amino acid residues and the calculated molecular weight is about 161 kD. BLAST analysis indicated that the gene may belong to pleiotropic drug resistance （PDR） sub-family, and hence designated as TaPDR1 （Triticum aestivum pleiotropic drug resistance）. TaPDR1 was located on chromosome 5A of wheat using nullisomic-tetrasomic lines of Chinese Spring. TaPDR1 was up-regulated by induction of both DON and F. graminearum. Expression patterns of TaPDR1 were different in wild-type Wangshuibai and the fast-neutron induced Wangshuibai mutant lacking FHB1, a major QTL of FHB resistance and DON resistance in chromosome arm 3BS. These results suggested that TaPDR1 might be a candidate gene responsible for DON accumulation resistance. The expression profile showed that TaPDR1 expression was neither induced by hormones typically involved in biotic stress, such as JA and SA, nor by abiotic stresses, such as heat, cold, wounding and NaCI. However, TaPDR1 expression was regulated by Al^3＋ and [Ca^2＋], indicating that [Ca^2＋]1 might mediate the signal of TaPDR1 expression.     

用激光光声检测技术研究乙醛对苹果乙烯产量的影响

激光光声检测技术是有极高的检测灵敏度，并能够无侵入地、连续地监测微量气体浓度随时间的变化，用基于ＣＯ２激光器的腔内光声光谱仪研究忆树苹果乙烯产量的影响，证实了乙醛蒸气对苹果组织乙烯生物合成的阻碍作用，实验发现，苹果果皮及果肉组织对惭醛有很高的扩散阻力，乙醛处理完整苹果没有明显的效果，但处理去皮的苹果切片时对乙烯合成有明显的阻止作用，研究结果对苹果贮前的短时间无氧处理方法提供了实验依据。     

河套蜜瓜Cucumis melo CV Hetau成熟期中某些生理生化过程的变化

对河套密瓜成熟过程中生长速率、呼吸强度以及内源乙烯含量变化的研究表明：该种甜瓜属呼吸跃变型果实，从开花到成熟约为３８ｄ；呼吸跃交出现之前乙烯开始产生，其含量升高先于呼吸跃变．可溶性精含量在成熟过程中不断增高，在呼吸峰值时达到最大，淀粉含量出现一个由积累到降解的变化．此外也研究了瓜硬度、色泽和可溶性固形物的变化．     

臭氧水处理对红富士苹果在两种贮藏温度下 果实品质的影响

研究不同浓度臭氧水处理对红富士苹果在两种贮藏温度下的贮藏品质和生理的影响.分别采用0.3,0.6,0.9,1.2 mg/L的臭氧水处理红富士苹果,然后分别贮藏于13℃和0℃,定期测定贮藏温度下的果实的生理和品质指标.臭氧处理对苹果果实呼吸强度及乙烯的释放均有一定的抑制作用;延缓苹果果实的可溶性固形物含量、可滴定酸含量及硬度的下降,减少果实水分的损失;0℃贮藏的保鲜效果普遍优于13℃贮藏条件.臭氧处理对红富士苹果具有较好的保鲜效果,0.9 mg/L臭氧水处理的效果优于其他各组,且0.9 mg/L臭氧水结合低温的贮藏方式更有利于红富士苹果贮藏品质的保持.     

鲜切南瓜组织中乙烯产生及1-氨基环丙烷-1-羧酸(ACC)氧化酶活性的变化

本文以鲜切南瓜为实验材料,研究了切割伤害诱导、外源乙烯和降冰片二烯NBD(2,5-norbonadien)处理对乙烯的产生和ACC氧化酶活性的影响.结果表明,南瓜切割和外源乙烯处理都可诱导乙烯产生和ACC氧化酶活性急速上升,达到高峰后逐渐下降并恢复到接近原来的水平.NBD可有效抑制由伤害诱导的ACC氧化酶活性的增加,进而抑制了乙烯的产生.但当外源乙烯和NBD存在时,ACC氧化酶活性不受NBD抑制,其活性明显增加,然后逐渐下降,72h后恢复到接近原来水平,而乙烯的产生显示了与对照相似的显著增加的结果.实验结果认为切割机械伤害和乙烯处理都可通过诱导ACC氧化酶活性的提高而使乙烯的产生显著增加,NBD对ACC氧化酶的抑制作用可被乙烯所解除.     

成熟猕猴桃果实不同组织中ACC氧化酶基因的表达差异研究

了猕猴桃“海沃特”成熟果实各组织中ACC氧化酶基因的转录水平和翻译水平对乙烯生成的影响，结果表明：（1）乙烯生成具有组织差异性；（2）ACC氧化酶基因mRNA的表达在各组中水平相似；（3）ACC氧化酶的表达水平具有显著的组织差异性及时间顺序性，因此，ACC氧化酶的瑶达水平的高低可能是导致乙烯生成的组织差异性的主要原因之一。     

氰丙氨酸合酶在乙烯诱导杨树耐盐中的作用

【目的】探讨木本植物线粒体β-氰丙氨酸合成酶在乙烯诱导的交替呼吸氧化酶(AOX)途径对盐胁迫响应中的作用。【方法】选取盐胁迫下‘南林895’杨叶片,利用HPLC测定氨基环丙烷羧酸(ACC,乙烯前体)含量,实时荧光定量PCR分析基因表达,比色法测定半胱氨酸水平。【结果】盐胁迫使杨树幼苗叶片ACC积累,乙烯合成相关酶(ACS7和ACO3)、氰丙氨酸合酶(CYS C1),以及腈水解酶(NIT4)等基因显著上调表达,同时伴随着线粒体交替呼吸氧化酶(AOX1b)基因的上调和细胞色素c氧化酶(COX6b)基因下调表达。水杨基氧肟酸(SHAM)预处理导致AOX1b基因表达被抑制,电解质渗透率(EL)和丙二醛(MDA)含量上升,但不影响CYS C1表达。而乙烯合成抑制剂氨基氧乙酸(AOA)了抑制CYS C1和盐胁迫诱导的AOX1b基因的表达,并增加EL和MDA含量。此外,AOA恢复盐胁迫减少的半胱氨酸含量,而SHAM和抗霉素A(AA)均无此效应。【结论】杨树叶片CYS C1参与了乙烯激发的耐盐响应,但乙烯诱导的交替呼吸氧化酶(AOX)并未位于CYS C1上游而发挥作用。     

不同苹果品种抗寒性的研究

为了检测适宜加工苹果不同品种在石河子的抗寒适应性,筛选适宜在石河子地区大面积推广的加工苹果品种,采用试验室模拟田间低温环境方法对8个苹果品种1年生枝条分别在4、-15、-20、-25、-30、-35℃条件下进行预处理,并对其相对电导率并计算临界半致死温度(LT40)、可溶性蛋白、脯氨酸及丙二醛含量进行研究。结果显示,经过不同梯度低温处理后,寒富、国光的萌芽率显著较高;相对电导率显著较低;国光丙二醛含量显著性较低;寒富的脯氨酸含量与可溶性蛋白含量显著较高。结论:苹果枝条经过低温胁迫后,寒富、国光、甜格力在石河子地区抗寒能力较好。     

高温敏核不育水稻幼苗生长发育的热激反应

高温敏感核不育水稻培矮 64S 在热激处理后, 幼苗叶蛋白质组分与其可育条件下( 22℃ )相比, 表现出明显差异:在分子量为 97. 4~ 14. 4 KD 范围内共表现出 6 条蛋白质区带的变化, 其中 4 条新增加, 2 条减弱. 在热激条件下, 与常规稻湘晚籼 2号相比较, 培矮 64S 亦表现出了 7条蛋白质区带的变化.这些蛋白质组分的改变必然引起体内某些酶系统的变化,而影响幼苗的正常生长发育, 使幼苗生长受到明显抑制.     

重组Pichia酵母(Muts)发酵过渡阶段关键酶活分析

重组Pichia酵母表达系统的发酵存在从利用甘油为碳源生长到利用甲醇为碳源表达外源蛋白的发酵表达过渡阶段。Pichia酵母过渡阶段碳源代谢途径关键酶酶活分析表明：甲醇诱导3h AOX2酶活为0,4h时突然增加到0．05U,继续诱导,酶活缓慢增加;在过渡阶段甲醛脱氢酶和6-P-葡萄糖脱氢酶分别增加了6．1倍、2．5倍,而丙酮酸脱氢酶和异柠檬酸脱氢酶分别下降为原酶活的29．4％及16．4％,表明甲醇诱导后甲醇完全氧化代谢途径得到强化,而糖酵解途径和三羧酸循环途径代谢作用减弱。