苹果果实中细胞质型苹果酸酶基因（NADP-ME）的克隆与表达分析

【研究目的】从苹果果实中克隆细胞质型NADP-苹果酸酶(NADP-ME)基因,并且从基因转录水平上研究该基因与苹果果实酸度的关系。【方法】利用RT-PCR和RACE技术克隆NADP-ME基因,半定量RT-PCR分析该基因在高酸和低酸个体不同发育时期果实中的转录情况,并进行原核表达分析。【结果】成功克隆了细胞质型NADP-ME基因的cDNA全长序列,命名为Mal-ME(GenBank注册号:DQ280492);该基因表达一个约66kD的蛋白;半定量RT-PCR分析表明:不同发育时期的果实中Mal-ME的转录与苹果酸含量呈负相关,即随着Mal-ME转录水平的增强果实含酸量下降。【结论】成功从苹果果实中克隆了细胞质型Mal-ME基因,Mal-ME在果实发育过程中对苹果酸的积累起负调控作用。     

Cytoplasmic relatedness of apple landraces and cultivars: a molecular analysis

Summary Mitochondrial DNA (mtDNA) diversity has been determined in apple landraces and cultivars, based upon restriction fragment length polymorphisms (RFLPs) for several mitochondrial genes. Our analysis includes three cultivars, Golden Delicious, McIntosh and Delicious, which represent the various patterns of mtDNA polymorphisms previously described (Ishikawa et al., 1992). A total of five RFLPs were detected, allowing classification of the apple genotypes into four cytoplasmic groups: GroupI, Golden Delicious-type; GroupII, McIntosh-type; GroupIII, Delicious-type; and Group IV, Dolgo Crab-type. European landraces and cultivars were assigned to Groups I, II, and III, while chinese crab apples were placed in either Group III or IV.     

Effects of Cinnamon Acid on Respiratory Rate and Its Related Enzymes Activity in Roots of Seedlings of Malus hupehensis Rehd

This paper studied the effects of cinnamon acid treatments on the respiratory rate and related enzymes activity in the seedling roots of Malus hupehensis Rehd.It would provide information for understanding the mechanisms of inhibition damage caused by continuous cultivation of apple tree.20 mL of solution containing different concentrations of cinnamon acid was added into container with the tested seedlings.After treatment,the samples were taken periodically and the respiratory rates were measured by OXY-LAB oxygen electrodes under 25°C stable temperature and then the activities of related enzymes were measured.The rates of total respiration and other 2 pathways [tricarboxylic acid cycle （TCA） and pentose phosphate pathway （PPP）] appeared initially an increasing treads and late （on the 3rd d） began to decline.However,they again appeared an increase trend at the end period,on the contrast,the respiratory rate of embden-meyer- hot-parnas （EMP） pathway appeared a stead decline tread but it had a recover on the last day.The respiratory rate of total and 3 pathways were decreased under 125 mg kg-1 （soil）.The dynamic trends of the enzymes activities of pyrophosphate-dependent phosphofructokinase （PFK）,glucose-6-phosphate dehydrogenase （G-6-PDH） and malate dehydrogenase （MDH） showed similarly.In conclusion,treatments of certain concentration of cinnamon acid would inhibit the respiratory rate and related enzymes activity of roots of M.hupehensis Rehd.And the inhibition degrees were positively related with concentration of cinnamon acid treatments.     

来安花红品种特性及发展状况

JIN Le-zhong(Laian Agricultural Technology Extension Center,Laian,Anhui 239200)     

Soil fumigation and compost amendment alter soil microbial community composition but do not improve tree growth or yield in an apple replant site

Apple replant disease (ARD) is a disease complex that reduces survival, growth and yield of replanted trees, and is often encountered in establishing new orchards on old sites. Methyl bromide (MB) has been the fumigant used most widely to control ARD, but alternatives to MB and cultural methods of control are needed. In this experiment, we evaluated the response of soil microbial communities and tree growth and yield to three pre-plant soil treatments (compost amendment, soil treatment with a broad-spectrum fumigant, and untreated controls), and use of five clonal rootstock genotypes (M.7, M.26, CG.6210, G.30 and G.16), in an apple replant site in Ithaca, New York. Polymerase chain reaction (PCR)—denaturing gradient gel electrophoresis (DGGE) analysis was used to assess changes in the community composition of bacteria and fungi in the bulk soil 8, 10, 18 and 22 months after trees were replanted. PCR-DGGE was also used to compare the community composition of bacteria, fungi and pseudomonads in untreated rhizosphere soil of the five rootstock genotypes 31 months after planting. Tree caliper and extension growth were measured annually in November from 2002 to 2004. Apple yield data were recorded in 2004, the first fruiting year after planting. Trees on CG.6210 rootstocks had the most growth and highest yield, while trees on M.26 rootstocks had the least growth and lowest yield. Tree growth and yield were not affected by pre-plant soil treatment except for lateral extension growth, which was longer in trees growing in compost-treated soil in 2003 as compared to those in the fumigation treatment. Bulk soil bacterial PCR-DGGE fingerprints differed strongly among the different soil treatments 1 year after their application, with the fingerprints derived from each pre-plant soil treatment clustering separately in a hierarchical cluster analysis. However, the differences in bacterial communities between the soil treatments diminished during the second year after planting. Soil fungal communities converged more rapidly than bacterial communities, with no discernable pattern related to pre-plant soil treatments 10 months after replanting. Changes in bulk soil bacterial and fungal communities in response to soil treatments had no obvious correlation with tree performance. On the other hand, rootstock genotypes modified their rhizosphere environments which differed significantly in their bacterial, pseudomonad, fungal and oomycete communities. Cluster analysis of PCR-DGGE fingerprints of fungal and pseudomonad rhizosphere community DNA revealed two distinct clusters. For both analyses, soil sampled from the rhizosphere of the two higher yielding rootstock genotypes clustered together, while the lower yielding rootstock genotypes also clustered together. These results suggest that the fungal and pseudomonad communities that have developed in the rhizosphere of the different rootstock genotypes may be one factor influencing tree growth and yield at this apple replant site.     

苹果SUPERMAN类蛋白家族的基因克隆与生物信息学分析

SUPERMAN 类基因是植物生长发育中的重要转录因子。为了揭示苹果 SUPERMAN 类蛋白家族的生理基因功能,本研究以富士苹果(Malus domestica Borkh.cv. Fuji)为试材,利用 RT-PCR 方法和锚定PCR 方法克隆了 MdLIFa(GenBank 登录号 HM370493)和启动子序列,利用特异引物 PCR 方法克隆了苹果基因组中其余 11 个 SUPERMAN 类基因。12 个基因命名为 MdLIFa/MdLIFb (GenBank 登录号:HQ260429)、MdSUP1a (HQ418477)/MdSUP1b (HQ418478)、MdSUP4 (HQ418475)/MdSUP12 (HQ418476)、MdSUP5a (HQ418469)/MdSUP13b (HQ418470)、MdSUP9a (HQ418473)/MdSUP9b (HQ418474) 和 MdSUP5b(HQ418471)/MdSUP13a(HQ418472),分别位于 11/3、1/1、4/12、5/13、9/ 未知和 5/13 号染色体,每对基因间的同源性均在86%以上。通过基因枪法轰击洋葱(Allium cepa)表皮细胞,基因瞬时表达证明 MdLIFa 定位于细胞核内。MdLIFa 基因启动子序列中含有多个参与花粉发育、根特异性表达、与光诱导和 Dof 单锌指基因家族作用的调控元件等。系统进化分析表明,MdLIFa/b、MdSUP9a/9b 和 MdSUP5a/13b、以及MdSUP1a/b、MdSUP5b/MdSUP13a和 MdSUP4/12 分别聚类为一组,参与植株的形态建成和花器官发育。苹果 12 个 SUPERMAN 类基因成对高度同源,具有生物学功能的多样性,为深入的基因鉴定和遗传转化提供有用信息。     

Effects of Plant and Soil Factors on Growth of Young Apple and Blackcurrant Plants

Abstract

Shoot growth as affected by plant size at planting, peat application in the planting hole, pruning, drip-irrigation and, for apple, specific apple replant disease (SARD) was investigated in apple (cv. Elstar) and blackcurrant (cvs Ben Lomond and Ben Nevis), using a multifactorial design. In the first year after planting, peat application and drip-irrigation enhanced shoot growth in both species, with a further interactional increase when both treatments were applied. The extension growth of apple, in contrast to blackcurrant, was positively related to plant size, and the strength of the relationship was distinctly improved when growth conditions were good. Pruning had only small effects on total shoot growth, with a trend towards an increase when growth conditions were poor.     

晋西黄土区幼龄苹果+花生间作地土壤水分的时空分布特征

选取晋西黄土区具有典型代表性的幼龄苹果(Malus pumila)+花生(Arachis hypogaea)间作地作为研究对象,在花生生长季的不同时间对苹果+花生间作地和对照花生单作地的土壤水分进行定位监测,研究苹果+花生间作系统间作界面上土壤水分的时空分布特征和水分效应,及其对花生生长状况和产量的影响。结果表明:1)间作地土壤平均水分含量生长季逐月变化显著,水分耗用最大的时间为7月;2)在垂直方向上,土壤含水量随着土壤深度的增加而增加,在水平方向上,土壤含水量最低值出现在距离果树行最近的区域,并随着与树行距离的增加而增加;3)间作地种间土壤水分耗用量最大的区域为靠近苹果树的表层土壤;4)在当前树龄下,苹果+花生间作系统相对于花生单作土壤水分在整体上表现为负效应,对花生的产量产生了不利的影响,并限制了果农间作系统生态效益和经济效益的进一步提高。建议采取适当的调控和管理措施缓解种间水分竞争并提高花生的产量。     

The effects of apple dwarfing rootstocks on leaf nutrient element composition in stoolbed production

The nutritional status of 22 apple (Malus domestica Borkh.) dwarfing rootstocks, with varied degrees of dwarfing, was evaluated during stoolbed production in 1983 and 1984. Rootstock effects were observed for all leaf nutrient elements studied except Cu. Except for EMLA 27, leaf N levels were generally highest in dwarfing EMLA and Polish series and lowest on vigorous MAC and EMLA rootstocks. Relatively few significant differences were found in leaf P and K content among rootstocks. Foliar Ca and Mg concentrations were generally highest on Polish and Budagovski rootstocks and lowest on EMLA 111. Leaf B levels on vigorous and semi‐dwarfing EMLA rootstocks and Fe levels in MAC, Polish, and Budagovski series were considerably higher than on other rootstocks. Although significant differences were found in leaf Mn and Zn among rootstocks, no specific effect for rootstock vigor was observed for either element. Positive correlations between leaf Ca and Mg occurred in 17 rootstocks while negative correlations between N and Fe were observed in 21 rootstocks.     

Start-Timing for Calcium Chloride Spray Programs Influences Fruit Calcium and Bitter Pit in ‘Braeburn’ and ‘Honeycrisp’ Apples

Early-season sprays of calcium chloride (CaCl₂) have been reported to sometimes be more effective than later-season sprays for controlling bitter pit in apple (Malus × domestica Borkh.). We conducted a two-year study examining the influence of start-timing of CaCl₂ spray programs on fruit Ca concentrations and at-harvest bitter pit incidence in ‘Braeburn’ and ‘Honeycrisp’ apples in Washington State, USA. Six biweekly sprays of CaCl₂ were applied starting in mid-May (early-start), mid-June (mid-start, the normal commercial start timing), or mid-July (late-start) 2002 and 2003. Although leaf marginal necrosis occurred in all of the CaCl₂-treated plots, no spray damage on fruit was observed. All of the CaCl₂ spray treatments consistently reduced bitter pit incidence relative to the unsprayed control treatment and usually but not always increased the Ca concentration of subdermal cortical tissue at harvest. The early and mid-start spray programs produced lower bitter pit incidence than the late-start program in only one of four situations. Fruit Ca concentrations in the control and early-start spray program did not differ. Fruit Ca concentrations in the mid- and late-start spray programs were not different, and often were higher than that of the early-start spray program. Bitter pit incidence for each cultivar was inversely related to fruit Ca concentration. Substituting cortical Ca/potassium (K) or Ca/magnesium (Mg) concentration ratios for Ca concentration failed to substantially improve and usually degraded the correlations with bitter pit incidence. The results confirm that starting CaCl₂ spray programs in June sometimes is more effective at controlling bitter pit that delaying the start of the spray program until July. The later season sprays tend to be more effective at increasing fruit Ca concentration. There appears to be no detectable advantage for starting spray programs earlier than June. Applying CaCl₂ sprays throughout the growing season, starting sometime in June, appears to be the most economical and effective spray management practice for maximizing fruit Ca and minimizing risk of bitter pit development.