平邑甜茶两类新生根诱导应用及其生物信息数据库构建
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摘要
根系是果树的根本，调控根系结构与功能是果园土壤管理的关键。课题组对蓬莱市曲受彭同志果园能够实现连年亩产量在1万公斤左右高产现象的多年调查，发现该果园土壤有机质含量高，吸收根量大，表层环境稳定；以此为背景，本文通过对大田和盆栽条件下的根系环境改良诱导研究，探讨新生根起源与根系构型、两类新生根对植株地上部生长发育的影响及其栽培培养的诱导应用研究；同时，利用差异表达谱测序技术，对两类新生根基因表达谱差异进行了分析，并通过对苹果生物信息学数据搜集，建立了苹果生物信息数据库平台。主要研究结果如下：
一、根组及两类新生根来源
通过不同介质的栽培试验发现，有机质能够诱导多分枝根系构型，并将此类根系构型命名为“根组”。根组是生长根在有机质土壤条件下诱导发生的多分枝根系构型，其发生部位主要是生长根侧生的多级分支。不同树龄和介质条件下，根组发生有显著影响，幼苗吸收根量少，且细弱；结果期大树有机质中吸收根粗壮。根组可以阶段性大量发生吸收根，是吸收根发生的主要母体。
解剖结构观察发现，‘平邑甜茶’新生根维管柱存在二原型、三原型、四原型和五原型的差异。生长根与吸收根均属于侧根发生方式，起源于中柱鞘细胞。吸收根的母根多为二原型，而生长根母根多为四原型和五原型。
二、两类新生根对植株生长发育的影响
不同介质的盆栽试验表明：改善效果顺序为混合处理>生物碳>有机肥单独处理>粘壤土处理。5%生物炭肥与10%有机肥混合使用叶绿素含量和净光合速率分别提高了15.55%和57.64%；植株生物量提高了46.36%。
局部改良的盆栽试验结果表明：中心区域为粘壤土，外层水氮诱导有效地提高了株高和生物量。以中层水氮诱导处理植株长势最好，株高比对照增加23.99%，主干粗度增加9.85%，生物量增加21.90%。不同改良位置对叶片质量和枝条贮藏营养无显著影响。
十年生‘富士’田间局部改良第二年，树体外部、中部及内部叶片净光合速率分别提高了27.23%、31.95%和46.39%；叶绿素含量分别提高了19.03%、12.52%和12.06%。外围延长枝春梢可溶性糖及淀粉分别提高了49%和33.5%，秋稍分别提高了28.30%和27.27%。顶芽和腋芽鳞片数均比对照多1-2片，叶原数增加1-2片。增产效果明显，单株增产18.38kg。
因此，吸收根能够提高叶片质量，防止叶片衰老，增加枝条贮藏营养，调控树势，提高花芽质量，提高产量。而生长根可有利于主轴延长、提高树势。
三、两类根系的栽培诱导
不同介质的栽培试验发现，5%生物炭肥单独使用及与有机肥混合使用增加了根系表面积、根尖数量和细根吸收面积比例。其中5%生物炭肥单独使用使根系表面积和根尖数量比对照增加了4.47倍和10.78倍，细根吸收面积比例为24.99%。生物炭肥与有机肥混合处理根系表面积和根尖数量增加5.03倍和14.83倍，细根吸收面积比例22.10%。
差异介质的水氮诱导试验发现，水氮能有效诱导根系再生，不同介质条件下，水氮诱导的根系差异显著。蛭石栽培下的水氮诱导根系以细根为主，新生生长根细弱；有机质栽培下水氮诱导的生长根和吸收根数量均明显增加，生长根粗长；有机质栽培下的水氮诱导，即能促进生长根的产生，又能诱导吸收根大量发生。土壤盆栽下的水氮能够诱导大量生长根产生，围绕盆壁，形成“界面效应”。河沙栽培下水氮诱导的生长根延长，少分支。
层次诱导试验发现，土壤区域水氮处理能够有效诱导根系延伸，长度为72.6cm，是介质诱导根系的2.62倍；土壤中水氮诱导根系以生长根吸收为主，细根吸收仅占1.28%。
不同层次的差异诱导‘平邑甜茶’盆栽试验发现，中心区域为生物炭肥与有机肥的改良介质时，外侧无根系外沿诱导效应，根系集中在中层和上层，其中上层根系占到62.09%以上。中心区域为土壤时，外沿介质区域产生大量细根，根系集中在介质中层，根系密度均高于5.10条/cm3，土壤区域水氮诱导产生大量长根，以中层水氮诱导效果最佳，盆栽‘平邑甜茶’可诱导出5-7条吸收根。故“外围诱导，中层水氮”。
十年生‘富士’田间局部改良当年，40×40×40cm的同等空间内，细根增加了45条，粗根增加8条；改造空间内根系总表面积增加6.76倍，根尖数量增加11.14倍。改良后第二年，细根增加了137条，增加粗根26条；根系表面积和根尖数量分别增加13.60倍和19.63倍。
因此，环境差异可导致明显的根系构型差异，通过生物炭肥、有机肥和化肥的不同肥料效果，可实现对‘平邑甜茶’根系的差异诱导，故“断根水氮诱导更新，基质诱导养根壮树”，且“局部改良，整体效果”。
四、“局部优化、三层节水”果园管理制度
总结群众经验和过去苹果根系研究成果及局部效应的特点，课题组提出“表层保护，局部优化，中层微补，下层贮水”节水节肥养根壮树的土壤管理体系，简称“三层节水”。“三层节水”处理后细根和粗根分别增加121%和112.5%；短枝比例和短枝质量增加；叶片厚度显著提高，栅栏组织细胞排列紧密，叶绿素分别提高7.1%和12.8%；顶芽和腋芽及叶原鳞片数增加1-2片；叶片POD含量增加，MDA含量降低，缓解了叶片膜脂过氧化进程；产量和品质提高。
五、利用差异表达谱测序技术，对两类新生根中差异表达基因进行了研究，从5,913,044和6,236,711条表达标签中筛选到了1,646个差异基因。广泛参与次级代谢途径、植物与病原互作、植物激素信号转导、苯丙烷代谢途径、核糖体及淀粉和蔗糖代谢及黄酮类代谢等。多数基因具有细胞内功能定位（82.7%）、催化活性（71.1%）、离子绑定（59.2%）及跨膜转运（24.5%）等功能定位。并验证了磷转运蛋白、钾离子通道、锌铁离子转运蛋白等在吸收根高表达。
六、本文通过搜集了广泛的网络资源，以结构域、基因家族和基因功能为单位对苹果基因进行了广泛的分类和功能注释，并构建了生物信息数据库。同时，数据库还包含广泛的基因信息，包括CDS及蛋白质序列、染色体定位、基因结构、GO分类、蛋白质结构域分析、Interpro分类、表达谱、MiRNARNA及相关文献等；此外，数据库还提供了用户交互平台，研究人员可提交基因信息及相关文献等参与苹果基因注释。为了方便浏览与使用，构建了公开的网络数据库平台，网址是：http://www.applegene.org/。
Root is the basis of the fruit trees, root structure and function regulation is the key oforchard soil management. The orchard per-acre yields achieved around10,000kg by QuShoupeng in PengLai in successive years. According to the research survey for many years,we found that the orchard had high content of soil organic horizons, large number ofabsorption roots, and stable surface circumstance. In this paper, the roots origin and formation,two types root function, and inducing application research of cultivation was discussed infield and pot cultivation. Meanwhile, expression profiles of sequencing technologydifferences were applied to compare the differential genes of the two types of new roots. Theapple biological information data base flat was set up by collecting the apple biologicalinformation data. The results were as follows:
1. Root group and the origin of two types of new roots
Through the results of phenotype observation, we put forward a new concept about theroot configuration,‘root group’, which is the branched root system configuration which isinduced by growing root under the condition of organic soil. Multilevel branches of the lateralgrowing root are the sites where root group forms.The difference in root group with tree ageand root mediums is significant. The seedlings absorbing root is less and thin, while strongand long-lived roots are formed in the region of organic matter during fruit stage. In differentstages, root group can induce a large number of absorbing root, and is the main parent root ofabsorbing root.
The root anatomy showed that the primary xylem of new roots was divided into diarch,triarch, tetrarchy and pentarch. Growing root and absorbing root are all originated in thepericycle cells, blonging to the lateral root development way. Most of absorbing root iscoming from diarch, while the growing root is coming from tetrarchy or pentarch.
2. The function of two types of new roots
The potted experiment of one year old Malus hupehensis in different mediums indicatedthat,5%biochar fertilizer mixed with10%organic fertilizer treatment made the chlorophyllcontent and net photosynthetic rate increase by15.55%and57.64%respectively, and plantbiomass increase by46.36%.
The result of local optimization of potted two years old Malus hupehensis in differentposition indicated that, the treatment of clay loam in central area and water-nitrogen inducingin outer area effectively increased the plant’s height and biomass. Plants grow best under thewater-nitrogen inducing treatment in the middle area, whose plant height, stem coarseness andbiomass increased by23.99%,9.85%and21.90%, respectively. Local optimization had nosignificant effect on leaf quality and storage nutrion of branches.
Local optimization in field made the photosynthetic rate of internal, central and externalleaf net increase by27.23%,31.95%and27.23%respectively; Chlorophyll content increaseby19.03%,12.52%and12.06%respectively. Solubility sugar and starch in spring shootsincrease by49%and33.5%, respectively. Solubility sugar and starch in autumn shootsincrease by28.30%and27.27%, respectively. The number of apical and axillary bud scales is1-2pieces more than control. Yield per plant in the second year significantly increasedfor18.38kg.
So, absorbing roots can improve leaf quality, prevent the senescence of leaf, increasebranches’ storage nutrition, regulate the growth of autumn shoot, improve the quality offlower bud, and increase fruit yield.
3. The inducing cultivation of two types of new roots
The study of potted experiment with different media indicated that,5%biochar treatmentmade root surface area and root tips number increase4.47times and10.78times than thecontrol. Biochar fertilizer mixed with organic fertilizer treatment made root surface area androot tips number increased by5.03times and14.83times. Biochar fertilizer used alone orcombined with organic fertilizer significantly increased fine root absorption area ratio, for24.99%and22.10%of total root surface area, respectively.
Water-nitrogen induced roots regeneration. Water-nitrogen induced most thin roots whenvermiculite as a growing medium, and the new growth roots were thin; the roots had morebranches when rganic matter as a growing medium, Water-nitrogen induced growing rootsand absorbing roots significantly increased, the absorbing roots were long and thick;
Water-nitrogen induced more growing roots and the absorbing roots when rganic matteras a growing medium. Water-nitrogen induced potted plants to form a large number ofgrowing roots, surrounded the inside of pots, which was called ‘surface interfacial effect’. Water-nitrogen induced the growing roots lengthened and had less branches when river sandsas a growing medium. So, ensuring the organic matter content and environmental stability inthe fertilizer region is the basis of reasonable induction of the growing roots and the absorbingroots. Layer induction experiments showed that water-nitrogen could induce rootslengthening to72.6cm, which was2.62times than the medium induction; water-nitrogeninduced more absorbing roots, and the thin roots were only1.28%.
The result of local optimization of potted two years old Malus hupehensis in differentposition showed that, no roots inducing along the outside and root was concentrated in themiddle and upper soil when the middle area was medium, the roots in upper mediumaccounted for more than62.09%. More thin roots formed outside when the central area wasclay loam, water-nitrogen induced more long roots, most roots formed in the mid-mediumarea, the average dendity of roots were more than5.10per cubic metres, and the spaceutilization efficiency was the best. Water-nitrogen inductive in the middle layer had the besteffect, which induced5to7long roots per cm3.
Local optimization in field made fine root number and thick root number increased by45and eight in the first year, respectively, made total surface area increased by6.76times, madethe number of root tips increase by11.14times. Local optimization in field made fine rootnumber, thick root number increase by137and26, respectively, made root surface and roottips number increased by13.60times and19.63times, respectively.
4. Local optimization and three layers of water-saving soil management system
Depending on the apple tree roots’ function and the inductive identity, the research groupput forward soil management system “protecting the top soil, optimizing local part,micro-irrigating the moderate soil, collecting water in the subsoil”, The three layers ofwater-saving treatment on ‘Fuji’ and ‘Gala’ showed that, the shoot and flowers quality wasimproved, the energy consumption was reduced, and the production significantly increased.
5. To gain insight into the transcriptome dynamics between the two types of new roots,genome-wide gene expression profiling was conducted by Solexa sequencing. More than fivemillion tags were generated from two type new roots, including1,646DEGs were identified,which participated in biosynthesis of secondary metabolites, plant-pathogen interaction, planthormone signal transduction, phenylpropanoid biosynthesis, ribosome and flavonoid biosynthesis. Most genes were identifid in the cell part function (82.7%), catalytic activity(71.1%), ion binding (59.2%) and membrane transcription (24.5%), etc. Next, the expressionpatterns of the ion transporting genes were assessed by quantitative real-time PCR, and theresults obtained showed general agreement with the Solexa analysis.
6. To analyze the function and evolution of different genes in root, we developed applegene function and gene family database (AppleGFDB) for collecting, storing, arranging, andintegrating functional genomics information of the apple. The AppleGFDB provides severallayers of information about the apple genes, including nucleotide and protein sequences,chromosomal locations, gene structures, and any publications related to these annotations. Tofurther analyze the functional genomics data of apple genes, the AppleGFDB was designed toenable users to easily retrieve information through a suite of interfaces, including geneontology, protein domain and InterPro. In addition, the database provides tools for analyzingthe expression profiles andmicroRNAs of the apple.Moreover, all of the analyzed andcollected data can be downloaded from the database. The database can also be accessed usinga convenient web server that supports a full-text search, a BLAST sequence search, anddatabase browsing. Furthermore, to facilitate cooperation among apple researchers,AppleGFDB is presented in a user-interactive platform, which provides userswith theopportunity to modify apple gene annotations and submit publication information for relatedgenes. AppleGFDB is available at http://www.applegene.org.
一、根组及两类新生根来源
通过不同介质的栽培试验发现，有机质能够诱导多分枝根系构型，并将此类根系构型命名为“根组”。根组是生长根在有机质土壤条件下诱导发生的多分枝根系构型，其发生部位主要是生长根侧生的多级分支。不同树龄和介质条件下，根组发生有显著影响，幼苗吸收根量少，且细弱；结果期大树有机质中吸收根粗壮。根组可以阶段性大量发生吸收根，是吸收根发生的主要母体。
解剖结构观察发现，‘平邑甜茶’新生根维管柱存在二原型、三原型、四原型和五原型的差异。生长根与吸收根均属于侧根发生方式，起源于中柱鞘细胞。吸收根的母根多为二原型，而生长根母根多为四原型和五原型。
二、两类新生根对植株生长发育的影响
不同介质的盆栽试验表明：改善效果顺序为混合处理>生物碳>有机肥单独处理>粘壤土处理。5%生物炭肥与10%有机肥混合使用叶绿素含量和净光合速率分别提高了15.55%和57.64%；植株生物量提高了46.36%。
局部改良的盆栽试验结果表明：中心区域为粘壤土，外层水氮诱导有效地提高了株高和生物量。以中层水氮诱导处理植株长势最好，株高比对照增加23.99%，主干粗度增加9.85%，生物量增加21.90%。不同改良位置对叶片质量和枝条贮藏营养无显著影响。
十年生‘富士’田间局部改良第二年，树体外部、中部及内部叶片净光合速率分别提高了27.23%、31.95%和46.39%；叶绿素含量分别提高了19.03%、12.52%和12.06%。外围延长枝春梢可溶性糖及淀粉分别提高了49%和33.5%，秋稍分别提高了28.30%和27.27%。顶芽和腋芽鳞片数均比对照多1-2片，叶原数增加1-2片。增产效果明显，单株增产18.38kg。
因此，吸收根能够提高叶片质量，防止叶片衰老，增加枝条贮藏营养，调控树势，提高花芽质量，提高产量。而生长根可有利于主轴延长、提高树势。
三、两类根系的栽培诱导
不同介质的栽培试验发现，5%生物炭肥单独使用及与有机肥混合使用增加了根系表面积、根尖数量和细根吸收面积比例。其中5%生物炭肥单独使用使根系表面积和根尖数量比对照增加了4.47倍和10.78倍，细根吸收面积比例为24.99%。生物炭肥与有机肥混合处理根系表面积和根尖数量增加5.03倍和14.83倍，细根吸收面积比例22.10%。
差异介质的水氮诱导试验发现，水氮能有效诱导根系再生，不同介质条件下，水氮诱导的根系差异显著。蛭石栽培下的水氮诱导根系以细根为主，新生生长根细弱；有机质栽培下水氮诱导的生长根和吸收根数量均明显增加，生长根粗长；有机质栽培下的水氮诱导，即能促进生长根的产生，又能诱导吸收根大量发生。土壤盆栽下的水氮能够诱导大量生长根产生，围绕盆壁，形成“界面效应”。河沙栽培下水氮诱导的生长根延长，少分支。
层次诱导试验发现，土壤区域水氮处理能够有效诱导根系延伸，长度为72.6cm，是介质诱导根系的2.62倍；土壤中水氮诱导根系以生长根吸收为主，细根吸收仅占1.28%。
不同层次的差异诱导‘平邑甜茶’盆栽试验发现，中心区域为生物炭肥与有机肥的改良介质时，外侧无根系外沿诱导效应，根系集中在中层和上层，其中上层根系占到62.09%以上。中心区域为土壤时，外沿介质区域产生大量细根，根系集中在介质中层，根系密度均高于5.10条/cm3，土壤区域水氮诱导产生大量长根，以中层水氮诱导效果最佳，盆栽‘平邑甜茶’可诱导出5-7条吸收根。故“外围诱导，中层水氮”。
十年生‘富士’田间局部改良当年，40×40×40cm的同等空间内，细根增加了45条，粗根增加8条；改造空间内根系总表面积增加6.76倍，根尖数量增加11.14倍。改良后第二年，细根增加了137条，增加粗根26条；根系表面积和根尖数量分别增加13.60倍和19.63倍。
因此，环境差异可导致明显的根系构型差异，通过生物炭肥、有机肥和化肥的不同肥料效果，可实现对‘平邑甜茶’根系的差异诱导，故“断根水氮诱导更新，基质诱导养根壮树”，且“局部改良，整体效果”。
四、“局部优化、三层节水”果园管理制度
总结群众经验和过去苹果根系研究成果及局部效应的特点，课题组提出“表层保护，局部优化，中层微补，下层贮水”节水节肥养根壮树的土壤管理体系，简称“三层节水”。“三层节水”处理后细根和粗根分别增加121%和112.5%；短枝比例和短枝质量增加；叶片厚度显著提高，栅栏组织细胞排列紧密，叶绿素分别提高7.1%和12.8%；顶芽和腋芽及叶原鳞片数增加1-2片；叶片POD含量增加，MDA含量降低，缓解了叶片膜脂过氧化进程；产量和品质提高。
五、利用差异表达谱测序技术，对两类新生根中差异表达基因进行了研究，从5,913,044和6,236,711条表达标签中筛选到了1,646个差异基因。广泛参与次级代谢途径、植物与病原互作、植物激素信号转导、苯丙烷代谢途径、核糖体及淀粉和蔗糖代谢及黄酮类代谢等。多数基因具有细胞内功能定位（82.7%）、催化活性（71.1%）、离子绑定（59.2%）及跨膜转运（24.5%）等功能定位。并验证了磷转运蛋白、钾离子通道、锌铁离子转运蛋白等在吸收根高表达。
六、本文通过搜集了广泛的网络资源，以结构域、基因家族和基因功能为单位对苹果基因进行了广泛的分类和功能注释，并构建了生物信息数据库。同时，数据库还包含广泛的基因信息，包括CDS及蛋白质序列、染色体定位、基因结构、GO分类、蛋白质结构域分析、Interpro分类、表达谱、MiRNARNA及相关文献等；此外，数据库还提供了用户交互平台，研究人员可提交基因信息及相关文献等参与苹果基因注释。为了方便浏览与使用，构建了公开的网络数据库平台，网址是：http://www.applegene.org/。
Root is the basis of the fruit trees, root structure and function regulation is the key oforchard soil management. The orchard per-acre yields achieved around10,000kg by QuShoupeng in PengLai in successive years. According to the research survey for many years,we found that the orchard had high content of soil organic horizons, large number ofabsorption roots, and stable surface circumstance. In this paper, the roots origin and formation,two types root function, and inducing application research of cultivation was discussed infield and pot cultivation. Meanwhile, expression profiles of sequencing technologydifferences were applied to compare the differential genes of the two types of new roots. Theapple biological information data base flat was set up by collecting the apple biologicalinformation data. The results were as follows:
1. Root group and the origin of two types of new roots
Through the results of phenotype observation, we put forward a new concept about theroot configuration,‘root group’, which is the branched root system configuration which isinduced by growing root under the condition of organic soil. Multilevel branches of the lateralgrowing root are the sites where root group forms.The difference in root group with tree ageand root mediums is significant. The seedlings absorbing root is less and thin, while strongand long-lived roots are formed in the region of organic matter during fruit stage. In differentstages, root group can induce a large number of absorbing root, and is the main parent root ofabsorbing root.
The root anatomy showed that the primary xylem of new roots was divided into diarch,triarch, tetrarchy and pentarch. Growing root and absorbing root are all originated in thepericycle cells, blonging to the lateral root development way. Most of absorbing root iscoming from diarch, while the growing root is coming from tetrarchy or pentarch.
2. The function of two types of new roots
The potted experiment of one year old Malus hupehensis in different mediums indicatedthat,5%biochar fertilizer mixed with10%organic fertilizer treatment made the chlorophyllcontent and net photosynthetic rate increase by15.55%and57.64%respectively, and plantbiomass increase by46.36%.
The result of local optimization of potted two years old Malus hupehensis in differentposition indicated that, the treatment of clay loam in central area and water-nitrogen inducingin outer area effectively increased the plant’s height and biomass. Plants grow best under thewater-nitrogen inducing treatment in the middle area, whose plant height, stem coarseness andbiomass increased by23.99%,9.85%and21.90%, respectively. Local optimization had nosignificant effect on leaf quality and storage nutrion of branches.
Local optimization in field made the photosynthetic rate of internal, central and externalleaf net increase by27.23%,31.95%and27.23%respectively; Chlorophyll content increaseby19.03%,12.52%and12.06%respectively. Solubility sugar and starch in spring shootsincrease by49%and33.5%, respectively. Solubility sugar and starch in autumn shootsincrease by28.30%and27.27%, respectively. The number of apical and axillary bud scales is1-2pieces more than control. Yield per plant in the second year significantly increasedfor18.38kg.
So, absorbing roots can improve leaf quality, prevent the senescence of leaf, increasebranches’ storage nutrition, regulate the growth of autumn shoot, improve the quality offlower bud, and increase fruit yield.
3. The inducing cultivation of two types of new roots
The study of potted experiment with different media indicated that,5%biochar treatmentmade root surface area and root tips number increase4.47times and10.78times than thecontrol. Biochar fertilizer mixed with organic fertilizer treatment made root surface area androot tips number increased by5.03times and14.83times. Biochar fertilizer used alone orcombined with organic fertilizer significantly increased fine root absorption area ratio, for24.99%and22.10%of total root surface area, respectively.
Water-nitrogen induced roots regeneration. Water-nitrogen induced most thin roots whenvermiculite as a growing medium, and the new growth roots were thin; the roots had morebranches when rganic matter as a growing medium, Water-nitrogen induced growing rootsand absorbing roots significantly increased, the absorbing roots were long and thick;
Water-nitrogen induced more growing roots and the absorbing roots when rganic matteras a growing medium. Water-nitrogen induced potted plants to form a large number ofgrowing roots, surrounded the inside of pots, which was called ‘surface interfacial effect’. Water-nitrogen induced the growing roots lengthened and had less branches when river sandsas a growing medium. So, ensuring the organic matter content and environmental stability inthe fertilizer region is the basis of reasonable induction of the growing roots and the absorbingroots. Layer induction experiments showed that water-nitrogen could induce rootslengthening to72.6cm, which was2.62times than the medium induction; water-nitrogeninduced more absorbing roots, and the thin roots were only1.28%.
The result of local optimization of potted two years old Malus hupehensis in differentposition showed that, no roots inducing along the outside and root was concentrated in themiddle and upper soil when the middle area was medium, the roots in upper mediumaccounted for more than62.09%. More thin roots formed outside when the central area wasclay loam, water-nitrogen induced more long roots, most roots formed in the mid-mediumarea, the average dendity of roots were more than5.10per cubic metres, and the spaceutilization efficiency was the best. Water-nitrogen inductive in the middle layer had the besteffect, which induced5to7long roots per cm3.
Local optimization in field made fine root number and thick root number increased by45and eight in the first year, respectively, made total surface area increased by6.76times, madethe number of root tips increase by11.14times. Local optimization in field made fine rootnumber, thick root number increase by137and26, respectively, made root surface and roottips number increased by13.60times and19.63times, respectively.
4. Local optimization and three layers of water-saving soil management system
Depending on the apple tree roots’ function and the inductive identity, the research groupput forward soil management system “protecting the top soil, optimizing local part,micro-irrigating the moderate soil, collecting water in the subsoil”, The three layers ofwater-saving treatment on ‘Fuji’ and ‘Gala’ showed that, the shoot and flowers quality wasimproved, the energy consumption was reduced, and the production significantly increased.
5. To gain insight into the transcriptome dynamics between the two types of new roots,genome-wide gene expression profiling was conducted by Solexa sequencing. More than fivemillion tags were generated from two type new roots, including1,646DEGs were identified,which participated in biosynthesis of secondary metabolites, plant-pathogen interaction, planthormone signal transduction, phenylpropanoid biosynthesis, ribosome and flavonoid biosynthesis. Most genes were identifid in the cell part function (82.7%), catalytic activity(71.1%), ion binding (59.2%) and membrane transcription (24.5%), etc. Next, the expressionpatterns of the ion transporting genes were assessed by quantitative real-time PCR, and theresults obtained showed general agreement with the Solexa analysis.
6. To analyze the function and evolution of different genes in root, we developed applegene function and gene family database (AppleGFDB) for collecting, storing, arranging, andintegrating functional genomics information of the apple. The AppleGFDB provides severallayers of information about the apple genes, including nucleotide and protein sequences,chromosomal locations, gene structures, and any publications related to these annotations. Tofurther analyze the functional genomics data of apple genes, the AppleGFDB was designed toenable users to easily retrieve information through a suite of interfaces, including geneontology, protein domain and InterPro. In addition, the database provides tools for analyzingthe expression profiles andmicroRNAs of the apple.Moreover, all of the analyzed andcollected data can be downloaded from the database. The database can also be accessed usinga convenient web server that supports a full-text search, a BLAST sequence search, anddatabase browsing. Furthermore, to facilitate cooperation among apple researchers,AppleGFDB is presented in a user-interactive platform, which provides userswith theopportunity to modify apple gene annotations and submit publication information for relatedgenes. AppleGFDB is available at http://www.applegene.org.
引文
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