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。     

根层调控措施对甜玉米-黄瓜设施蔬菜轮作体系土壤硝态氮的影响

【目的】以甜玉米作为填闲作物,探讨不同的根层调控措施对消减土壤剖面累积硝态氮及下茬黄瓜生长的影响。【方法】在华北平原传统棚室蔬菜的休闲季种植甜玉米,针对甜玉米设置添加土壤调理剂和秸秆还田2种根层调控措施,以甜玉米传统种植作为对照,进行田间小区试验。试验于2008年5月至2011年5月进行,共3次甜玉米-黄瓜轮作,6季作物。每年6月初至9月底种植甜玉米,10月初至次年1月底扣棚育黄瓜苗,当年2月初种植黄瓜。在甜玉米季,共3个处理,随机排列,重复3次。小区面积为4 m×2 m,小区间隔0.3 m,区组之间布设1 m的保护行。【结果】甜玉米种植季,调理剂处理的玉米籽粒产量最高,2008、2009和2010年的产量分别为6.2、7.4和7.9 t·hm^-2;土壤调理剂和秸秆还田2种根层调控处理的甜玉米总吸氮量高于传统种植。秸秆还田和调理剂处理能够促进20-60 cm土层根系的生长发育,促使根系吸收更深层的土壤养分。2种根层调控措施均能降低土壤剖面NO₃^--N的累积,尤其对100-200 cm的作物根区NO₃^--N的消减能力更强,NO₃^--N消减趋势大致为：调理剂>秸秆还田>传统种植。3季黄瓜种植季,不同前茬处理的黄瓜产量、生物量和吸氮量差异均不显著;3季平均土壤NO₃^--N在0-200 cm土层的残留量为秸秆还田<调理剂<传统种植。3个轮作季后,传统种植、调理剂和秸秆还田处理在0-200 cm土层的氮素盈余量分别为1 911.6、1 966.3和1 930.2 kg·hm^-2,调理剂处理显著高于传统种植。【结论】在硝态氮高累积的设施土壤上,随着种植年限的增加,加入土壤调理剂和适当的秸秆还田对100-200 cm的作物根区土壤剖面NO₃^--N的消减能力更强。填闲作物种植第二年对下茬黄瓜土壤NO₃^--N的消减作用最为明显。土壤调理剂和秸秆还田措施能够显著提高甜玉米对土壤剖面NO₃^--N的消减能力,减缓土壤NO₃^--N 的淋失,提高经济效益。     

Histopathology of infection and colonization of Quercus ilex fine roots by Phytophthora cinnamomi

Quercus ilex is one of the European forest species most susceptible to root rot caused by the oomycete Phytophthora cinnamomi. This disease contributes to holm oak decline, a particularly serious problem in the ‘dehesas’ ecosystem of the southwestern Iberian Peninsula. This work describes the host–pathogen interaction of Q. ilex and P. cinnamomi, using new infection indices at the tissue level. Fine roots of 6‐month‐old saplings inoculated with P. cinnamomi were examined by light microscopy and a random pool of images was analysed in order to calculate different indices based on the measured area of pathogen structures. In the early stages of invasion, P. cinnamomi colonizes the apoplast and penetrates cortical cells with somatic structures. On reaching the parenchymatous tissues of the central cylinder, the pathogen develops different reproductive and survival structures inside the cells and then expands through the vascular system of the root. Some host responses were identified, such as cell wall thickening, accumulation of phenolic compounds in the middle lamella of sclerenchyma tissues, and mucilage secretion blocking vascular cells. New insights into the behaviour of P. cinnamomi inside fine roots are described. Host responses fail due to rapid expansion of the pathogen and a change in its behaviour from biotrophic to necrotrophic.     

The endophytic fungus Piriformospora indica protects wheat from fusarium crown rot disease in simulated UK autumn conditions

The root endophytic fungus Piriformospora indica (Sebacinacea) forms mutualistic symbioses with a broad range of host plants, increasing their biomass production and resistance to fungal pathogens. This study evaluated the effect of P. indica on fusarium crown rot disease of wheat, under in vitro and glasshouse conditions. Interaction of P. indica and Fusarium isolates under axenic culture conditions indicated no direct antagonistic activity of P. indica against Fusarium isolates. Seedlings of wheat were inoculated with P. indica and pathogenic Fusarium culmorum or F. graminearum and grown in sterilized soil‐free medium or in a non‐sterilized mix of soil and sand. Fusarium alone reduced emergence and led to visible browning and reduced root growth. Roots of seedlings in pots inoculated with both Fusarium isolates and P. indica were free of visible symptoms; seed emergence and root biomass were equivalent to the uninoculated. DNA was quantified by real‐time polymerase chain reaction (qPCR). The ratio of FusariumDNA to wheat DNA rose rapidly in the plants inoculated with Fusarium alone; isolates and species were not significantly different. Piriformospora indica inoculation reduced the ratio of Fusarium to host DNA in the root systems. The reduction increased with time. The ratio of P. indica to wheat DNA initially rose but then declined in root systems without Fusarium. With Fusarium, the ratio rose throughout the experiment. The absolute amount of FusariumDNA in root systems increased in the absence of P. indica but was static in plants co‐inoculated with P. indica.     

海南省无公害农产品产地建设中存在的问题及发展对策

研究了橡胶树小筒苗筒底的不同悬空培养高度对其接穗生长的影响,为橡胶树优良小筒苗的培育提供理论依据。结果表明：小筒苗的根系穿出情况与筒底悬空高度密切相关,不同悬空高度对小筒苗的长势也具有显著影响;穿根率高的小筒苗长势较好,但定植费工费时,成活率较低;培育小筒苗适宜的控根隔离层高度是4 cm或4 cm以上,此时其大田定植成活率相对较高。     

不同悬空培养高度对橡胶树小筒苗接穗生长的影响

研究了橡胶树小筒苗筒底的不同悬空培养高度对其接穗生长的影响,为橡胶树优良小筒苗的培育提供理论依据。结果表明：小筒苗的根系穿出情况与筒底悬空高度密切相关,不同悬空高度对小筒苗的长势也具有显著影响;穿根率高的小筒苗长势较好,但定植费工费时,成活率较低;培育小筒苗适宜的控根隔离层高度是4 cm或4 cm以上,此时其大田定植成活率相对较高。     

Floral traits and pollination system of Zygophyllum xanthoxylum in the managed and wild populations in an arid region of Northwest China

Zygophyllum xanthoxylum, which belongs to Sarcozygium of Zygophyllaceae, is one of the ecologically important species in Northwest China. In order to understand the pollination system of Z. xanthoxylum, we investigated the following characteristics of this species in the Urat Desert-grassland Research Station in western Inner Mongolia of China: flowering dynamics, pollen viability, pollen limitation, floral visitors and breeding system. The results showed that the flowering period and flowering peak were different between the wild and managed populations, being longer in the managed population. Z. xanthoxylum was pollen-limited, and pollen limitation was more intense in the wild population than in the managed population. Chalicodoma deserticola(Hymenoptera) was found to be the most frequent pollinator in the wild population, while Anthophora fulvitarsis(Hymenoptera) was the most frequent and effective visitor in the managed population. Out-crossing was dominant in the breeding system and self-pollination just played an assistant role to assure the reproduction of Z. xanthoxylum.     

Evidence of low levels of genetic diversity for the Phytophthora austrocedrae population in Patagonia,Argentina

Phytophthora austrocedrae is a recently discovered pathogen that causes severe mortality of Austrocedrus chilensis in Patagonia. The high level of susceptibility of the host tree, together with the distribution pattern of the pathogen, have led to the hypothesis that P. austrocedrae was introduced into Argentina. The aim of this study was to assess the population structure of P. austrocedrae isolates from Argentina in order to gain an understanding of the origin and spread of the pathogen. Genetic diversity was determined based on amplified fragment length polymorphisms (AFLPs). In total, 48 isolates of P. austrocedrae were obtained from infected A. chilensis trees, representing the geographical range of the host. Four primer combinations were used for the AFLP analysis. Of the 332 scored bands, 12% were polymorphic. Gene diversity (h) ranged from 0·01 to 0·03; the Shannon index (I) ranged from 0·01 to 0·04. A high degree of genetic similarity was observed among the isolates (pairwise S values = 0·958–1; 0·993 ± 0·009, mean ± SD). A frequency histogram showed that most of the isolate pairs were identical. Principal coordinate analysis using three‐dimensional plots did not group any of the isolates based on their geographical origin. The low genetic diversity (within and between sites) and absence of population structure linked to geographic origin, together with the aggressiveness of the pathogen and the disease progression pattern, suggest that P. austrocedrae might have been introduced into Argentina.     

Genetic diversity of the root‐knot nematode Meloidogyne ethiopica and development of a species‐specific SCAR marker for its diagnosis

Meloidogyne ethiopica is an important nematode pathogen causing serious economic damage to grapevine in Chile. In Brazil, M. ethiopica has been detected with low frequency in kiwifruit and other crops. The objectives of this study were to evaluate the intraspecific genetic variability of M. ethiopica isolates from Brazil and Chile using AFLP and RAPD markers and to develop a species‐specific SCAR‐PCR assay for its diagnosis. Fourteen isolates were obtained from different geographic regions or host plants. Three isolates of an undescribed Meloidogyne species and one isolate of M. ethiopica from Kenya were included in the analysis. The results showed a low level of diversity among the M. ethiopica isolates, regardless of their geographical distribution or host plant origin. The three isolates of Meloidogyne sp. showed a high homogeneity and clustered separately from M. ethiopica (100% bootstrap). RAPD screenings of M. ethiopica allowed the identification of a differential DNA fragment that was converted into a SCAR marker. Using genomic DNA from pooled nematodes as a template, PCR amplification with primers designed from this species‐specific SCAR produced a fragment of 350 bp in all 14 isolates of M. ethiopica tested, in contrast with other species tested. This primer pair also allowed successful amplification of DNA from single nematodes, either juveniles or females and when used in multiplex PCR reactions containing mixtures of other root‐knot nematode species, thus showing the sensitivity of the assay. Therefore, the method developed here has potential for application in routine diagnostic procedures.