Isolation of Trichoderma virens 6PS-2 and its effects on Fusarium proliferatum f. sp. Malus domestica MR5 related to apple replant disease in China

Apple replant disease(ARD) is primarily caused by biotic factors that seriously inhibits the development of apple industry. Therefore, the use of biological control measures to inhibit the main pathogens(such as Fusarium spp.) that cause ARD is of great significance to the sustainable development of the apple industry. Trichoderma virens 6PS-2, which exhibited antagonism toward a variety of pathogens, was screened from the rhizosphere soils of healthy apple trees(Malus robusta) in different replanted orchards in the Yantai and Zibo Cities, Shandong Province, China. Its fermentation extract inhibited the growth of pathogenic Fusarium proliferatum f. sp. Malus domestica MR5, which was proportional to the concentration. These substances also increased the hairy root volume and growth of Arabidopsis thaliana lateral roots. The phenotype of Malus hupehensis seedlings and microbial community structure in rhizosphere soils in greenhouse experiment using Highthroughput sequencing were analyzed, and the field experiment with grafted apple trees were used for further verification. Compared with the application of potato dextrose broth(PDB) medium, application of 6PS-2 spore suspension directly to replanted soils could improve the growth of M. hupehensis seedlings as well as the elongation of grafted apple trees. Concomitant decreases in the gene copy number of Fusarium and increases in the culturable bacteria/fungi were also observed in the greenhouse and field experiments. The abundance of Trichoderma,Bacillus, and Streptomyces increased significantly, but that of Fusarium, Pseudarthrobacter, and Humicola decreased. The content of esters, phenols,furans, and amino acids in root exudates of M. hupehensis seedlings increased, which significantly inhibited the multiplication of Fusarium, but was positively correlated with Bacillus and Trichoderma. In summary, T. virens 6PS-2 not only directly inhibits the activity of pathogenic Fusarium but also secrets secondary metabolites with antifungal and growth-promoting potential. In addition, 6PS-2 spore suspension can also promote the growth of plants to a certain extent, and change the soil microbial community structure of rhizosphere soils. It is believed that T. virens 6PS-2 has the potential for the alleviation of apple replant disease(ARD) in China.

MdDREB2A in apple is involved in the regulation of multiple abiotic stress responses

Abiotic stress has a serious effect on plant growth.The transcription factor DREB2A is a member of the AP2/ERF family,which is widely involved in abiotic stress response.However,the function of apple MdDREB2A has not been systematically investigated.In this study,MdDREB2A was isolated from the cultivar‘Royal Gala’.The open reading frame of MdDREB2A was 1197 bp in length and it encoded a protein of 398 amino acidswithmolecularweight of 43.8 kD.As a transcription factor,MdDREB2Awas located in the nucleus.qRT-PCR analysis showed that MdDREB2A was involved in responses to drought,salt,and ABA stresses.Under these stress treatments,the relative electrical conductivity,superoxide anion and malondialdehyde(MDA)in transgenic materials significantly decreased,and the content of proline increased in MdDREB2A transgenic plants,compared to the controls,indicating that MdDREB2A transgenic apple calli and transgenic Arabidopsis exhibited improved resistance to abiotic stress.This study introduces a candidate gene for the genetic improvement of crop resistance and reveals important function of MdDREB2A in the regulation of abiotic stress response.

Genome-wide identification,molecular evolution,and functional characterization of fructokinase gene family in apple reveal its role in improving salinity tolerance

Fructokinase(FRK)is a regulator of fructose signaling in plants and gateway proteins that catalyze the initial step in fructose metabolism through phosphorylation.Our previous study demonstrated that MdFRK2 protein exhibit not only high affinity for fructose,but also high enzymatic activity due to sorbitol.However,genome-wide identification of the MdFRK gene family and their evolutionary dynamics in apple are yet to be reported.A systematic genome-wide analysis in this study identified a total of nine MdFRK gene members,which could phylogenetically be clustered into seven groups.Chromosomal location and synteny analysis of MdFRKs revealed that their expansion in the apple genome is primarily driven by tandem and segmental duplication events.Divergent expression patterns of MdFRKs were observed in four source-sink tissues and at five different apple fruit developmental stages,which suggested their potential crucial roles in the apple fruit development and sugar accumulation.Reverse transcription-quantitative PCR(RT-qPCR)identified candidate NaCl or drought stress responsive MdFRKs,and transgenic apple plants overexpressing MdFRK2 exhibited considerably enhanced salinity tolerance.Our results will be useful for understanding the functions of MdFRKs in the regulation of apple fruit development and salt stress response.

Molecular cloning and functional identification of an apple flagellin receptor MdFLS2 gene

The leucine-rich repeat receptor kinase flagellin-sensing 2 gene(MdFLS2; Gene ID: MDP0000254112) was cloned from Royal Gala apple(Malus×domestica Borkh.). This gene contained a complete open reading frame of 3 474 bp that encoded 1 158 amino acids. The phylogenetic tree indicated that Prunus persica FLS2 exhibited the highest sequence similarity to MdFLS2. The PlantCare database suggests that the promoter sequence of MdFLS2 contains several typical cis-acting elements, including ethylene-, gibberellin-, salicylic acid-, and drought-responsive elements. Quantitative real-time PCR analysis showed that MdFLS2 was widely expressed in the different tissues of the apple and most highly expressed in the leaves. Furthermore, MdFLS2 was significantly induced by the flagellin elicitor peptide flg22. Treatment of the apple seedling leaves with flg22 resulted in an increase in leaf callose levels with increased treatment duration. An increase in the production of Oalong with the expression of disease-related genes was also observed. An oxidative burst was detected in the treated seedlings, but not in the control seedlings, indicating that flg22 had stimulated the expression of the MdFLS2 gene and its downstream target genes. Furthermore, the ectopic expression of MdFLS2 complemented the function of the Arabidopsis fls2 mutant and conferred enhanced flg22 tolerance to the transgenic Arabidopsis, suggesting that MdFLS2 acts as a positive regulator in the response to pathogens in apple.

MdDGK3-like as a negative regulator participates in ALA-induced PP2AC to promote stomatal opening in apple leaves

5-Aminolevulinic acid(ALA)can inhibit abscisic acid(ABA)-induced stomatal closure.However,the molecular mechanism is unclear.In this study,we found that ALA upregulated the MdPP2AC expression and PP2A activity in the apple(Malus domestica Borkh.cv.‘Fuji’)leaves.With the promoter of MdPP2AC as bait,a diacylglycerol kinase MdDGK3-like was selected by the Yeast One Hybrid(Y1H)from the cDNA library of the epidermis of apple leaves treated by exogenous ALA.Additional to binding the promoter of MdPP2AC,MdDGK3-like was found to inhibit the transcription activity of MdPP2AC promoter,while ALA significantly eliminated the role of MdDGK3-like.In tobacco leaves,MdDGK3-like was localized in the nucleus of stomatal guard cells.Therefore,MdDGK3-like might act as a transcription factor negatively regulating MdPP2AC expression and causing stomatal closure.To further identify MdDGK3-like functions,several transiently transgenic apple leaves(including overexpression and interference)were established.The results revealed that overexpression of MdDGK3-like promoted stomatal closure by increasing Ca^(2+)and H_(2)O_(2)and decreasing flavonol levels in the guard cells.Conversely,MdDGK3-like(i)led the stomatal opening with lower levels of Ca^(2+)and H_(2)O_(2)but higher flavonols.Based on these,we proposed a new hypothesis that ALA up-regulated MdPP2AC expression via negatively regulating the expression of MdDGK3-like to up-regulate PP2A expression and the enzyme activity,which improved the stomatal aperture.Since it was the first time that MdDGK3-like was showed to act as a transcription factor,the proposed model provided a new insight onto the mechanisms of ALA-induced stomatal opening.

Auxin response factor gene MdARF2 is involved in ABA signaling and salt stress response in apple

Auxin response factors(ARFs)play key roles throughout the whole process of plant growth and development,and mediate auxin response gene transcription by directly binding with auxin response elements(AuxREs).However,their functions in abiotic stresses are largely limited,especially in apples.Here,the auxin response factor gene MdARF2(HF41569)was cloned from apple cultivar‘Royal Gala’(Malus×domestica Borkh.).Phylogenetic analysis showed that ARF2 proteins are highly conserved among different species and MdARF2 is the closest relative to PpARF2 of Prunus persica,but they differ at the DNA level.MdARF2 contains three typical conserved domains including the B3 DNAbinding domain,Auxin_resp domain and AUX_IAA domain.The subcellular localization demonstrated that MdARF2 is localized in the nucleus.The three-dimensional structure prediction of the proteins showed that MdARF2 is highly similar with AtARF2,and they contain helices,folds,and random coils.The promoter of MdARF2 contains cis-acting elements which respond to various stresses,as well as environmental and hormonal signals.Expression analysis showed that MdARF2 is widely expressed in all tissues of apple,with the highest expression of MdARF2 in root.Functional analysis with a series of MdARF2 transgenic apple calli indicated that MdARF2 can reduce the sensitivity to ABA signaling and enhance salt tolerance in apple.In summary,the results of this research provide a new basis for studying the regulation of abiotic stresses by ARFs.

Apple发布DVD Studio Pro 2——简化专业DVD创作

Apple今天发布了DVD StudioPro2,这个经过重新设计的应用软件通过突破性用户界面简化了专业DVD创作,即日起通过苹果专卖店和苹果授权经销商对外发售、Apple设计的完全可定制的模板、创新的菜单编辑器、基于时间轴的轨道编辑和新的世界级MPEG-2编码器等新功能使DVDStudio Pro 2成为每一个创作专业人员必备的软件。 DVD Studio Pro

Biocontrol Efficacy of Three Antagonistic Yeasts Against Penicillium expansum in Harvested Apple Fruits

Three antagonistic yeasts, Trichosporon pullulans (Lindner.) Diddens and Lodder, Cryptococcus laurentii (Kuffer.) Skinner and Rhodotorula glutinis (Fresen.) F. C. Harrison, selected from over 50 yeasts on apple fruits, were investigated of their biocontrol efficacy against blue mould rot caused by Penicillium expansum Link in apple fruits. T. pullulans was identified to be a new yeast antagonist and was the most effective at inhibiting blue mould rot in the three yeasts. The effects of different concentrations of the three yeasts and addition Of nutrients, as well as combination with calcium on controlling blue mould rot in apples were presented in the paper. The results indicated that the higher the yeast concentrations, the more effective the yeasts on biocontrol activity. Addition of nutrients reduced the biocontrol capacity of the yeasts. Combination of 0.18 mol/L CaCl2 with C. laurentii cell suspensions could significantly enhance its effect to P. expansum in apple fruits, while the efficacy of calcium on biocontrol activity of T. pullulans or R. glutinis was not remarkable.

胃癌组织中APPL1和COX-2的表达及其与患者预后的关系

目的探讨胃癌组织中APPL1和COX-2蛋白表达情况、二者相关情况及其与患者生存期的关系。方法应用免疫组织化学SP法检测83例经手术切除石蜡包埋存档胃癌组织中APPL1和COX-2蛋白的表达情况。结果胃癌组织中APPL1和COX-2表达阳性率分别为60.2%(50/83)、62.7%(52/83)。APPL1的表达与胃癌的浸润深度、淋巴结转移、TNM分期、组织学类型差异有统计学意义(P<0.05);COX-2的表达与胃癌的浸润深度、淋巴结转移、TNM分期差异有统计学意义(P<0.05),而与组织学类型差异无统计学意义(P>0.05)。它们与肿瘤患者年龄、性别差异均无统计学意义(P>0.05)。相关分析结果显示APPL1与COX-2表达呈正相关(P<0.05)。生存曲线研究结果显示APPL1和COX-2表达与患者术后生存时间显著相关(P<0.05)。结论 APPL1和COX-2表达程度与胃癌的浸润深度、淋巴结转移、TNM分期密切相关,影响患者的生存时间。

‘长富2’ב金红’F1代苹果农艺性状的遗传变异分析

以‘长富2’ב金红’及其F1代为试材,对花期、叶片和果实性状进行遗传变异分析。结果表明,初花期以4月25日最多,开花持续天数以七八天居多。所有叶片性状、单粒重、果实横径、果实纵径、果型指数符合数量性状的遗传特点。叶片性状低于低亲率40.18%~83.04%,表现出明显的偏低遗传趋势;单果重、纵径和横径的平均值均小于亲本亲中值,且杂种优势率均为负值,表现出偏小的遗传倾向;酸低于低亲率48.21%,也表现出偏小的遗传倾向;果肉硬度表现出趋中的遗传趋势;可溶性固形物含量超高亲率为70.54%,表现出明显的超高亲遗传倾向。总之,杂交后代农艺性状呈现较广分离,有趋向于高糖和低酸的遗传趋势,同时叶片性状、单果重有趋向于低亲的遗传趋势。

施钾时期对长富2号苹果果实品质的影响

分别于采收前90、75、60、45、30 d对长富2号苹果园土壤进行施钾处理,探究采收前施钾时期对果实矿质营养、单果重、可溶性固形物含量、可滴定酸含量等的影响。结果表明,采收前施钾可提高长富2号果实氮、磷、钾含量,增加单果重、果形指数、花青苷含量、可溶性固形物含量,降低可滴定酸含量,促进着色。其中,采收前90 d施肥,单果重最大,可溶性固形物含量和钾含量最高;采收前75 d施肥,果实氮、磷含量最高,固酸比最大;采收前60 d施肥,果皮花青苷含量最高,a*值和CI值最大,果实着色好,果色偏红;采收前30 d施肥,单果重、果形指数、固酸比等与CK均无显著差异。综上,长富2号苹果采收前90 d至60 d增施钾肥对提高果实品质的效果最明显,为陕西省洛川县长富2号苹果园施用钾肥的适宜时期。

Extracellular and intracellular infection of Botryosphaeria dothidea and resistance mechanism in apple cells

Apple ring rot,which is caused by Botryosphaeria dothidea,severely affects apple production.The mechanisms employed in apple cells against B.dothidea remain unknown.In this research,the pathogen infection mode and the relationship between cell death and disease resistance in‘Fuji’/B.dothidea interaction pathosystem were investigated.By using transmission electron microscopy(TEM),our research showed that the pathogen infects apple cells both intracellularly and extracellularly.However,compared with that in immature fruit,the incidence of hyphae in the interior of mature apple fruit cells increased dramatically,suggesting that cell wall-mediated penetration resistance could be important in apple resistance against B.dothidea.TEM ultrastructural characterization identified the nuclear morphology of programmed cell death induction in both apple fruit and callus cells under B.dothidea infection.Overexpression of MdVDAC2(MDP0000271281),which encodes an outer-membrane localized anion channel protein in mitochondria,significantly promoted cell death under B.dothidea infection and simultaneously inhibited pathogen infection,suggesting that cell death represents a disease resistance mechanism in apple against B.dothidea infection.Furthermore,BdCatalase(KAF4307763),a cytochromeP450 family protein BdCYP52A4(KAF4300696),and subtilisin-domain containing proteinswere identified fromB.dothidea-secreted proteins,which suggested the potential involvement of active oxygen species and phytoalexins in combating B.dothidea infection and triggering or dampening apple resistance.Collectively,our research suggested that cell wall-mediated penetration resistance,programmed cell death machinery and microbial effector-interrelated signaling were among strategies recruited in apple to combat B.dothidea.The current research laid the foundation for further investigations into resistance mechanisms in apple.

Effect of Partial Root-Zone Irrigating Deuterium Oxide on the Properties of Water Transportation and Distribution in Young Apple Trees

Partial root-zone irrigation （PRI） has been proved to be an optimal water-saving irrigation technology, however, few studies were done on water transportation and distribution under PRI. The present study was performed to investigate the water transportation and distribution among the wet and dry root-zones and the shoot using deuterium water （D2O） in 1/4 root-zone PRI experiment. It also aimed to determine and analyze the D2O relative abundance within different types of roots and shoots. The results indicated that water could be transported from roots in wet root-zone to roots in dry root-zone and shoots within 2 h after irrigation. Water transportation in roots of wet-zone was carried out by absorbing root, 1-2 mm root, 2-5 mm root, and〉5 mm root progressively, while through a reverse process in three dry root-zones. In shoots, water was transported to trunk, central trunk, annual branches, shoot and leaf progressively. Thus in the young apple trees subjected to PRI, water was distributed ifrst in the roots, including the roots in the wet and dry root-zones, to satisfy the water need of roots itself, and then transported to the shoot within hours of irrigation.

S1B-3 Baicalin Modulates APPL2/Glucocorticoid Receptor Signaling Cascades,Promotes Neurogenesis,and Attenuates Emotional and Olfactory Dysfunctions in Chronic Corticosterone-Induced Depression

Olfactory dysfunction is accompanied with anxiety-and depressive-like behaviors.Impaired neurogenesis in hippocampus and subventricular zone(SVZ)-olfactory bulb(OB)contribute to anxiety-and depressive behaviors and olfactory dysfunctions.However,the underlying mechanisms remain unclear.Adaptor proteins containing the pleckstrin homology domain,phosphotyrosine binding domain,and leucine zipper motif(APPLs)are involved in regulating many biological activities.APPL2 showed the potentials to modulate cell growth,but whether APPL2 could affect adult neurogenesis and animal mood behaviors remains unknown.Herein,we tested the hypothesis that APPL2 could affect glucocorticoid receptor(GR)signaling and modulate hippocampal neurogenesis,contributing to depressive and anxiety behaviors.APPL2 Tg mice had enhanced GR phosphorylation under basic condition but had no different plasma corticosterone(CORT)level and GR phosphorylation under stress stimulation.APPL2 Tg mice had decreased hippocampal neurogenesis that was reversed by GR antagonist RU486.APPL2 Tg mice had impaired hippocampal neurogenesis and depressive and anxiety behaviors.We further identified the roles of APPL2 in olfactory functions.APPL2 Tg mice displayed higher GR activity and less neurogenesis at olfactory system with less olfactory sensitivity than WT mice,indicating that APPL2 could be a potential therapeutic target for depression and olfactory deficits.We then studied the effects of baicalin,a natural antioxidant,on modulating APPL2/GR signaling pathway and promoting neurogenesis for antidepressant and improving olfactory functions.Baicalin inhibited APPL2/GR signaling pathway and improved neurogenesis at SVZ,OB,and hippocampus in APPL2 Tg mice and chronic corticosterone-induced depression mouse model.Baicalin attenuated depressive-and anxietylike behaviors and improved olfactory functions in chronic depression mouse model and APPL2 Tg mice.In conclusion,APPL2 could be a novel therapeutic target for improving depressant related olfactory dysfunctions.Antioxidant therapy with baicalin could inhibit APPL2-mediated GR hyperactivity and promote neurogenesis,releasing depressive and anxiety symptoms and improving olfactory functions.

黑白酷睿 Apple MacBook华丽上市

只用90天的时间能创造什么？苹果电脑公司完成划时代的转变,推出所有新款MacBook系列。压轴的.就是采用英特尔“酷睿”双核处理器和华丽的全新13英寸光滑宽屏显示屏的最先进笔记本电脑。想想看，速度比小白快五倍，比小飞机快四倍是什么感觉？MaoBook采用最新的英特尔CoreDuo（酷睿）双核处理器以及包括一个667MHz前端总线和可扩展至2GB的667MHz DDR2 SDRAM内存除此之外．MacBook还相应地调整和升级了周边配置。其搭载的13英寸镜面宽屏显示屏有着极其艳丽的色彩表现力，同时在可视角度方面进一步优化，

苹果矮化砧木‘辽砧2号’选育

从1135系的助列涅特与M9杂交实生苗中选出优良苹果矮化砧木—辽砧2号(原代号80-1-6),经过22年田间观查和区试鉴定,其矮化性、早果性、丰产性、生根性与M26相近,适应性强于M26,以其为中间砧的富士树栽后6年进入盛果期,平均株产为23.5kg,同龄M26为17.2kg;8年生树冠积为4.07m3,同龄M26为4.58m3,树体存活率较M26高55%,平均每666.7m2产量1000kg以上;与基砧山定子和生产上主栽的富士系、元帅系、国光系等品种亲和性良好,以自根砧和中间砧方式利用均可。

不同矮化中间砧对‘长富2号’苹果生长特性、叶片生理及果实品质的影响

【目的】探究不同矮化中间砧对‘长富2号’苹果树体结构的组成、叶片抗氧化酶活性及果实品质的影响,为筛选适宜陇东地区密植的砧穗组合提供参考依据。【方法】以八棱海棠为基砧,SH40、M26、SH38、B9、M9、M9-T337、Y-1、GM256为中间砧,嫁接‘长富2号’为试验材料。【结果】8个砧穗组合的幼树矮化程度由强到弱依次为M9-T337>M9>M26>Y-1>GM256>SH40>SH38>B9。M9-T337、M26、M9树高和冠幅较其他砧穗组合明显减小,而主枝分支数量和干径横截面积显著高于其他组合。M9-T337、M26、M9的枝条生长量、枝条总量和长枝比例较其他砧穗组合明显减小,而短枝比例显著高于其他砧穗组合。M9-T337、M26和M9单果质量、硬度、果形指数和可溶性固形物含量显著高于其他砧穗组合,而可滴定酸含量明显减小。通过隶属函数法综合评价,得出各砧穗组合在生长势(致矮效果)、抗氧化酶活性和果实品质等方面的排序是M9-T337>M9>M26>SH38>SH40>Y-1>GM256>B9。【结论】陇东地区以八棱海棠为基砧,以M9-T337、M9和M26为中间砧,‘长富2号’作为接穗品种较佳,可考虑作为当地苹果栽培的首选砧穗组合。

不同产地长富2号苹果品质差异及其与地理坐标的相关性

对全国8个不同产地长富2号苹果的基本品质指标进行了测定,比较不同产地同一品种的品质差异,并分析地理坐标与品质指标的相关性及果实品质指标间的相关性.结果表明,不同产地长富2号品质差异较大,果形指数、果皮色度、总酸含量和固酸比差异明显.果实品质指标与地理坐标间相关性分析显示:果形指数和果实硬度分别与经度呈显著负相关;果实硬度和总酸含量分别与海拔呈极显著正相关;果实固酸比与海拔呈显著负相关;证明地理位置对同一品种苹果品质有影响.各品质指标间相关性分析显示:单果重分别与果形指数、可溶性固形物含量及总酸含量呈显著正相关;单果重、硬度、可溶性固形物含量均与总酸含量呈显著正相关.

钙肥对富士苹果品质及Ca^(2+)-ATPase活性影响的研究

以盛果期的矮化富士为材料,研究了不同钙肥对富士苹果品质和Ca2+-ATPase活性的影响.结果表明,喷钙后单果重增加,Vc含量提高,可溶性固形物和花青苷含量增大,而成熟果实的叶绿素和可滴定酸含量下降.不同钙肥效果顺序为:巨金钙>氨基酸钙>翠康钙宝>钙宝2000.钙肥对Ca2+-ATPase活性影响极为显著,其活性明显高于对照,不同钙肥间Ca2+-ATPase活性的变化趋势与果实品质的变化趋势相同.