Molecular cloning and functional characterization of MdPIN1 in apple

Auxin has been identified to play critical roles in regulating plant growth and development. The polar transport of auxin is regulated by auxin transporters. In the present study, an auxin efflux carrier gene MdPIN1 was cloned from Malus×domestic, Royal Gala, and introduced into wild-type Arabidopsis thaliana(Col-0). The transgenic plants exhibited the phenotype of inhibition of primary root(PR) elongation and increased lateral root(LR) number in compared with Col-0. Overexpression of Md PIN1 affected auxin transport, and enhanced phototropic responses and geotropism reaction, whereas had no significant difference in the auxin biosynthesis. These findings suggest that the Md PIN1 gene plays a vital role in auxin transport and root development.     

槲皮素促进拟南芥生长素极性运输的研究(英文)

Study on the role of quercentin in polar auxin transportation. Arabidopsis was cultured on medium supplemented with quercetin to observe the growth of hypocotyls,14C-IAA transport assays were conducted to measure the auxin transport activity. The results showed that Arabidopsis mutant aux1 which had been deficient in auxin influx transportion obviously recovered the ability after cultured on the medium with quercetin. The polar auxin transport was promoted by the addition of quercetin. These results indicated that quercetin could promote polar auxin transport in vivo.     

Quercetin Promotes Auxin Transport in Arabidopsis thaliana

Study on the role of quercentin in polar auxin transportation. Arabidopsis was cultured on medium supplemented with quercetin to observe the growth of hypocotyls,14C-IAA transport assays were conducted to measure the auxin transport activity. The results showed that Arabidopsis mutant aux1 which had been deficient in auxin influx transportion obviously recovered the ability after cultured on the medium with quercetin. The polar auxin transport was promoted by the addition of quercetin. These results indicated that quercetin could promote polar auxin transport in vivo.     

Effect of Phytohormones on Adventitious Bud Differentiation from Bulb Scales of Oriental Lily Test-tube Plantlets

[Objective] The purpose of this study is to investigate the effects of differ- ent phytohormones on the adventitious bud differentiation of oriental lily. [Method] The bulb scales of the test-tube plantlets of Tiber, Rodina and Constanta were cul- tured in media supplemented with different cytokinin and auxin at different concen- tration, and then the adventitious buds in each treatment were calculated. [Result] Cytokinins had different influence on the adventitious bud differentiation of the three oriental lily cultivars. Among them, 6-BA had the best effect to induce the adventi- tious bud differentiation from bulb scales of Tiber and Rodina, but there was some difference in the optimal concentration. KT had the best effect to induce the adven- titious bud differentiation of Constanta. The auxins had little influence on the quality of the adventitious bud of the three oriental lily cultivars, but caused some difference in differentiation coefficients. [Conclusion] The most suitable media for the adventi- tious bud differentiation from bulb scales in vitro of Tiber, Rodina and Constanta were MS＋0.2 mg/L 6-BA＋0.2 mg/L 2,4-D, MS＋I.0 mg/L 6-BA＋0.2 mg/L IAA, MS＋ 1.0 mg/L KT＋0.5 mg/L 2,4-D, respectively.     

Dormancy Characteristics of Malus ‘Snowdrift’ and Effects of Spraying 6-benzyladenine on Its Germination and Flowering in Long-day Treatment

[Objective] The dormancy characteristics of Malus ‘Snowdrift’ in long-day treatment were studied, and 6-BA was used to break the dormancy, with the aim to achieve the purpose of flowering in autumn. [Method] The new shoots of ‘Snowdrift’ in long-day treatment were conducted with hydroponics to investigate their dormancy time. And cytokinin 6-benzyladenine(6-BA)was used to treat the dormant shoots, to investigate the budding, flowering and flower bud differentiation. [Result] The shoots in long-day treatment entered endodormancy after August 7~(th) and the shoots in the natural daylight entered endodormancy before July 18~(th). In long-day treatment, 116 buds, 198 flowers were observed after 6-BA spraying. [Conclusion] The optimum concentration of 6-BA was 300 mg/L. And the flower bud differentiation of ‘Snowdrift’ in long-day treatment was faster than those in natural daylight after 6-BA spraying.     

生长素介导下三唑醇在大豆植株内的传导性(英文)

[Objective] This study was to investigate the mobility of IAA-triadimenol and NAA-triadimenol in soybean seedlings. [Method] Soybean seedlings were treated by spraying the solution of conjugated compounds onto their leaves; and chromatography was employed to measure the contents of IAA-triadimenol and NAA-triadimenol in different parts of these soybean seedlings, through which to represent the mobility of the conjugates. [Result] Both the triadimenol and NAA-triadimenol could not transport basipetally; whereas IAA-triademenol was ambimobile. When sprayed with 0.5 mmol/L IAA-triadimenol, as much as 1.87 μg/(g·FW) IAA-triadimenol was detected in the roots of soybean seedlings 12 h later, which was higher than that in the stem [0.68 μg/(g·FW)]; while that decreased to 0.80 μg/(g·FW) in the seedlings sprayed with 0.5 mmol/L IAA-triadimenol and the same concentration of IAA. [Conclusion] IAA moiety of the conjugate could enhance the transport and accumulation of the fungicide towards the root via the IAA carriers.     

SORPTION AND TRANSFORMATIONS OF ARSENITE AND ARSENATE IN SOILS AND THEIR RELATION TO RICE GROWTH

In the present paper,a series of laboratory,pot and field experiments was made to study the sorption and transformation of arsenite As(Ⅲ)and arsenate As(Ⅴboth in soils derived from 14 different parent materials and in various soil components as well as their relations between the sorption and transformation and rice growth,to elucidate the mechanisms of soil chemistry involved in the sorption and transformation of As(Ⅲ)and As(Ⅴ)by soils and soil components.These comprehensive studies have provided some effective measures of protecting and reversing arsenic pollution in the soil environment.The results obtained are summarised as follows     

Histological Study on the Polyovular Folliclular Development of Chinese Prematural Lubei White Goat

The present study was conducted to investigate the histological characteristics of Polyovular follicles in Chinese Lubei white female goats with age 2-5 months. The results suggested that the healthy antral follicles with twin-oocyte could be observed. There were many multi-oocyte phenomena in primordial and primary follicles with a proportion of nearly 75% in primordial follicles. Primordial follicle was in the outer of cortex in ovaries and most of them remained active. The nest of multi-nuclear primordial follicle was developing into primordial follicle with 1 to 3 nuclei.     

ROBUST REPETITIVE CONTROL FOR IMPROVING RATE SMOOTHNESS OF TEST TURNTABLE

A robust repetitive control scheme is used to improve the rate smoothness of a brushless DC motor (BLDCM) driven test turntable. The method synthesizes variable structure control (VSC) laws and repetitive control (RC) laws in a complementary manner. The VSC strategy can stabilize the system and suppress uncertainties, such as the aperiodic disturbance and noises, while RC strategy can eliminate the periodic rate fluctuation in a steady state. The convergence of the repetitive learning process is also guaranteed by VSC. A general nonlinear system model is discussed. The model can be considered as an extension of BLDCMs. The stability and asymptotic position tracking performance are validated by using Lyapunov functions. Simulation results show the effectiveness of the proposed approach for improving the rate smoothness.     

槲皮素促进拟南芥生长素极性运输的研究

用含有外源槲皮素的培养基种植拟南芥，探讨槲皮素对拟南芥生长素极性运输的影响。     

Quercetin Promotes Auxin Transport in Arabidopsis thaliana

Study on the role of quercentin in polar auxin transportation. Arabidopsis was cultured on medium supplemented with quercetin to observe the growth of hypocotyls, 14C-IAA transport assays were conducted to measure the auxin transport activity. The results showed that Arabidopsis mutant auxl which had been deficient in auxin influx transportion obviously recovered the ability after cultured on the medium with quercetin. The polar auxin transport was promoted by the addition of quercetin. These results indicated that quereetin could promote polar auxin transport in vivo.     

拟南芥液泡分拣蛋白AtVPS25调控植物生长素响应的功能分析

【目的】鉴定生长素胁迫条件下纯合突变体vps25的表型,获得拟南芥液泡分选蛋白AtVPS25的互作蛋白,并分析AtVPS25和其互作蛋白在生长素响应过程中的功能及其分子机制。【方法】根据“三引物法”鉴定突变体;通过观察拟南芥vps25突变体在外加生长素的培养基上的表型,鉴定AtVPS25的功能;以AtVPS25为诱饵蛋白,采用泛素分离系统筛选在拟南芥中与其互作的蛋白;利用酵母互作试验和双分子荧光互补试验（BiFC）验证AtVPS25与AtAIR12（for Auxin-Induced in Root cultures）的互作关系;鉴定AtVPS25和AtAIR12蛋白在植物细胞中的定位情况;采用Real-time PCR方法,分析在生长素处理条件下,部分生长素运输相关基因在拟南芥vps25突变体中的表达变化。【结果】Real-time PCR结果显示,10 μmol•L-1 IAA处理条件下,在野生型拟南芥（WT）中,AtVPS25的表达量随着胁迫时间的增长而增高,并在12 h达到最高,约为0 h的40倍,证明AtVPS25受生长素处理的诱导表达。利用泛素分离系统筛库获得AtVPS25的互作蛋白AtAIR12、AtVPS25与AtAIR12全长蛋白序列的酵母双杂交试验证明AtVPS25与AtAIR12互作。亚细胞定位试验证明AtVPS25定位在细胞膜和细胞质中,AtAIR12定位在细胞膜及叶绿体膜上。BiFC（双分子荧光互补）试验结果显示,AtVPS25蛋白与AtAIR12蛋白互作,并且互作位点在细胞膜和细胞质中。突变体鉴定获得纯合突变体vps25。vps25在0.1 mg•L-1 IAA条件下生长,表现为主根伸长受到抑制,并且相对同一条件下的WT的主根长度差异极显著（P＜0.01）,而同时侧根数无明显差异,这与已报道的air12-1突变体在生长素处理条件下的表型相似。10 μmol•L-1 IAA处理时,在WT背景条件下,AtAIR12的表达量对IAA响应明显,并且随着胁迫时间的增长而增高,在12 h时达到最高,约为0 h的80倍,证明10 μmol•L-1 IAA处理条件下,WT中AtAIR12表达量的变化趋势与AtVPS25完全相同。同时在vps25突变体背景条件下,AtAIR12的表达相对于WT受到抑制,在0-24 h表达量无明显变化。此外,在vps25突变体背景条件下,生长素输出载体基因AtPIN2相对于WT中表达量降低,生长素输入载体基因AtLAX2相对于WT中表达量提高。【结论】拟南芥液泡分拣蛋白基因AtVPS25受IAA诱导表达,参与调控植物主根的发育,AtVPS25可以与生长素响应蛋白AtAIR12在细胞质和细胞膜上互作,AtVPS25调控部分生长素相关基因的表达,AtVPS25通过调控这些下游基因的表达影响生长素在根部的响应。AtVPS25与AtAIR12的调控机制需要进一步深入研究。     

花生生长素输入载体基因PNAUX1的克隆及表达模式分析

生长素作为重要的植物激素,在调节植物生长发育中起重要作用。其所具有的极性运输特性,主要通过运输载体进行调控。本研究从花生未成熟种子cDNA文库中筛选到一个可能的生长素输入载体AUX/LAX类基因,命名为PNAUX1,通过RT-PCR对其表达模式分析显示,该基因在花生各组织中为组成型表达;同时,将PCR扩增获得的目的片段与植物双元表达载体pCAMBIA1301连接,构建了正义和反义载体转化DR5∷GUS拟南芥,并获得转基因植株。     

A role for flavin monooxygenase-like enzymes in auxin biosynthesis

Although auxin is known to regulate many processes in plant development and has been studied for over a century, the mechanisms whereby plants produce it have remained elusive. Here we report the characterization of a dominant Arabidopsis mutant, yucca, which contains elevated levels of free auxin. YUCCA encodes a flavin monooxygenase-like enzyme and belongs to a family that includes at least nine other homologous Arabidopsis genes, a subset of which appears to have redundant functions. Results from tryptophan analog feeding experiments and biochemical assays indicate that YUCCA catalyzes hydroxylation of the amino group of tryptamine, a rate-limiting step in tryptophan-dependent auxin biosynthesis.     

生长素在烟草表皮毛中的极性运输及对细胞伸长的影响

利用生长素极性运输转运蛋白PIN1与增强绿色荧光蛋白EGFP的融合，对PIN1进行了荧光标记，并以烟草表皮毛为模式，开展了PIN1对生长素极性运输及对细胞伸长生长影响的研究。采用DNA重组技术，将融合标记基因EGFP–PIN1置于拟南芥表皮毛特异表达基因GL2的启动子调控下，构建成含GL2pro::EGFP–PIN1的Ti质粒，以根癌农杆菌叶盘共培转化法将重组标记基因转化至烟草WS38中，筛选鉴定出多株转基因烟草。通过对这些转基因烟草表皮毛进行显微荧光观察，结果发现，标记的绿色荧光信号集中分布在表皮毛细胞的间隔区，表现为明显的极性分布现象。用生长素极性运输抑制剂三碘苯甲酸(TIBA)处理后，表皮毛的伸长生长受到抑制，细胞中荧光的分布极性减弱。说明生长素在烟草表皮毛中的极性分布对烟草表皮毛伸长起关键作用，抑制生长素的极性运输不只抑制表皮毛的细胞伸长，同时还影响到生长素极性运输蛋白PIN的极性分布。     

IAA对水稻根毛形成与水通道蛋白基因表达关系的研究

【目的】探讨气培条件下,IAA极性运输对水稻根毛形成及水通道蛋白基因表达的影响。【方法】以超表达OsPIN1a的转基因水稻和野生型水稻为研究材料,当种子根长为0.5-1.0 cm时,分别用不同浓度IAA、IAA和生长素极性运输输出载体抑制剂、IAA和生长素输入载体抑制剂复合处理的琼脂块贴在根尖一侧,黑暗条件下培养12 h后观察根毛生长情况,测定根毛的密度、长度及根毛形成后根尖部分相对含水量,并用激光共聚焦显微镜观察根尖及根毛内的OsPIN1a-GFP亚细胞定位,再通过半定量RT-PCR检测根毛形成前后水通道蛋白基因表达。【结果】（1）在0-5.0 mg·L^-1 IAA浓度范围内,随IAA浓度增加,根毛长度和密度也增加,当浓度超过5.0 mg·L^-1时,根毛长度和密度不再增加,根的生长受到显著抑制,2.5 mg·L^-1 IAA诱导效果最好;（2）只有根尖分生区IAA处理后,才能诱导新的根毛区形成根毛,而其他区域处理后无法诱导根毛形成;（3）根毛形成不受低浓度的生长素极性运输输出载体抑制剂TIBA、NPA和IAA复合处理的影响,高浓度TIBA、NPA和IAA复合处理则显著抑制根毛形成及根的生长;（4）低浓度生长素极性运输输入载体抑制剂CHPAA和IAA复合处理能显著抑制根毛形成和根的生长;（5）IAA不但能诱导根毛形成,也能诱导多种水通道蛋白基因表达,提高根尖相对含水量;（6）根尖及根毛内的OsPIN1a基因的表达和OsPIN1a-GFP含量均受IAA诱导,并在根毛细胞内大量表达。【结论】在气培条件下,根毛形成需要IAA诱导,而IAA诱导根毛形成过程需要生长素极性运输输入和输出载体蛋白的参与,其中输入过程对根毛的形成影响最大。IAA极性运输输入或输出载体蛋白特异性抑制剂处理显著降低根毛形成和根的生长,因此OsPIN1a在根毛形成中起着重要作用,同时根毛水通道蛋白基因表达量增加,提高了根尖相对含水量,减缓了气培条件下水稻根尖的水分胁迫。     

AXR4 is required for localization of the auxin influx facilitator AUX1

The AUX1 and PIN auxin influx and efflux facilitators are key regulators of root growth and development. For root gravitropism to occur, AUX1 and PIN2 must transport auxin via the lateral root cap to elongating epidermal cells. Genetic studies suggest that AXR4 functions in the same pathway as AUX1. Here we show that AXR4 is a previously unidentified accessory protein of the endoplasmic reticulum (ER) that regulates localization of AUX1 but not of PIN proteins. Loss of AXR4 resulted in abnormal accumulation of AUX1 in the ER of epidermal cells, indicating that the axr4 agravitropic phenotype is caused by defective AUX1 trafficking in the root epidermis.