CN110903367A - 转录因子在增加苹果枝条开张角度中的应用 - Google Patents
转录因子在增加苹果枝条开张角度中的应用 Download PDFInfo
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- CN110903367A CN110903367A CN201911236458.6A CN201911236458A CN110903367A CN 110903367 A CN110903367 A CN 110903367A CN 201911236458 A CN201911236458 A CN 201911236458A CN 110903367 A CN110903367 A CN 110903367A
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Abstract
本发明属于生物技术和植物基因工程技术领域，公开了一种转录因子在增加苹果枝条开张角度中的应用，本发明通过转基因手段，发现苹果中一个重要的转录因子MdMYB88/124，过表达之后，显著增加了苹果的枝条开张角度，果树开张角度变大，对于果树生产意义重大，增加通风透光，提高产量等，代替人工拉枝，有利于节省果园管理的成本。用量角器进行测量，没有转基因的嘎啦苹果枝条的平均角度为54.25度，MdMYB124过表达株系#1，#2，#19开张角度枝条平均开张角度为79度，74.9度，81.8度，MdMYB88的过表达株系#23平均开张角度为83.6度。
Description
技术领域
本发明属于生物技术和植物基因工程技术领域，尤其涉及一种转录因子在增加苹果枝条开张角度中的应用。
背景技术
目前，最接近的现有技术：开张角度大的株型可减少叶片间的相互遮挡，使其均匀接受光照，从而提高叶片光合效率，使叶片含碳量增加。对果树叶片营养物质含量与果实品质各指标进行相关及回归分析结果表明，果树枝条经不同拉枝角度处理后叶片碳素营养物质含量与果实单果重、果形指数、花色素含量、硬度、vC含量、总糖和蛋白质含量均呈正相关，与果实总酸含量呈负相关。因此，通过改良枝条和叶片开张角度可以提升果实品质。油菜分枝角度，通过QTL发现27个基因作为候选基因，包括了生长素响应基因，为破译油菜分枝角的遗传控制提供了依据。人为拉枝处理后，枝条分枝角度变大且生长方向发生改变，导致枝条上侧的GA和IAA含量显著上升，而下侧GA和IAA含量显著降低，且同一枝条上侧GA、IAA含量显著高于下侧，从而验证在枝条生长发育过程中，GA和IAA对枝条分枝角度具有重要影响。
综上所述，现有技术存在的问题是：现有技术通过人工和药剂，材料等管理增加了成本，技术实施难度较大。
解决上述技术问题的难度：激素双面作用，处理用量小起不到作用，用量大，则起反作用。拉枝时间和拉枝角度大小需要专业果树从业者进行。MYB88转基因材料，为培育开张角度大的苹果品种提供接穗，或者杂交亲本。并且为研究苹果开张角度增大提供实验材料。
解决上述技术问题的意义：本发明简化了管理，降低了成本。
发明内容
针对现有技术存在的问题，本发明提供了一种转录因子及表达载体和在增加果树枝条开张角度中的应用。
本发明是这样实现的，一种增加果树枝条开张角度的转录因子，其特征在于，所述增加果树枝条开张角度的转录因子MdMYB88/124，序列为：SEQ ID NO：1-SEQ ID NO：4。
本发明的另一目的在于提供一种由所述增加果树枝条开张角度的转录因子构建的表达载体，所述表达载体：pCAMBIA2300,pGWB414，pGWB415。
本发明的另一目的在于提供一种所述转录因子在增加果树枝条开张角度中的应用。
综上所述，本发明的优点及积极效果为：本发明通过转基因手段，发现苹果中一个重要的转录因子MdMYB88/124，过表达之后，显著增加了苹果的枝条开张角度，果树开张角度变大，对于果树生产意义重大，增加通风透光，提高产量等，代替人工拉枝，有利于节省果园管理的成本。用量角器进行测量，没有转基因的嘎啦苹果枝条的平均角度为54.25度，MdMYB124过表达株系#1，#2，#19开张角度枝条平均开张角度为79度，74.9度，81.8度，MdMYB88的过表达株系#23平均开张角度为83.6度。沉默株系与嘎啦的枝条夹角没有差异。并且测量叶片夹角，发现MdMYB88过表达株系叶柄夹角变大。
本发明通过增大果树枝条开张角度影响结实的机理，在20世纪五六十年代已研究的比较清楚，幼树拉枝可促进树冠扩大和提早成形，大树拉枝可改善树体的通风透光条件，促进果树内膛照光，提高光能利用率，从而促进果实着色和提高产量。近年的研究发现，外施植物激素也可以起到增大植物分枝夹角的作用。MdMYB88是在调控植物生长发育，非生物胁迫逆境中起到重要作用的基因。在观察MdMYB88转基因苹果材料时发现，MdMYB88的过表达株系幼苗叶夹角和两年生枝条开张角度均变大对于果树生产意义重大，增加通风透光，提高产量等，代替人工拉枝，有利于节省果园管理的成本。
本发明通过对MdMYB88转基因株系开张角度增大的功能进行研究，对实际生产和新品种培育起到重要作用。
附图说明
图1是本发明实施例提供的嘎啦苹果与转基因枝条夹角示意图；
图中：(a)GL-3；(b)MdMYB124 OE#1；(c)MdMYB124 OE#2；(d)MdMYB124 OE#19；(e)MdMYB88 RNAi#25；(f)MdMYB88 RNAi#24。
图2是本发明实施例提供的嘎啦苹果与转基因枝条夹角柱状图。
图3是本发明实施例提供的叶柄夹角示意图；
图中：(a)GL-3；(b)MdMYB88 OE。
图4是本发明实施例提供的MdMYB88转基因植物和GL-3的叶角示意图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
下面结合附图对本发明的应用原理作详细的描述。
本发明实施例提供的转录因子的序列为：MdMYB88的核苷酸序列为SEQ ID NO：1；MdMYB88的氨基酸序列为SEQ ID NO：2；MdMYB124的核苷酸序列为SEQ ID NO：3；MdMYB124的核苷酸序列为SEQ ID NO：4。MdMYB88的氨基酸序列为SEQ ID NO：2MdMYB88(XP_008389496.1)和MdMYB124(XP_008339925.1)。根据蔷薇科基因组网站(https://www.rosaceae.org/；GDR)提供的苹果基因组结构注释，发现苹果MdMYB88(MDP0000165523)，MdMYB124(MDP0000210970)基因全长分别为1620bp和1572bp，而NCBI(https://www.ncbi.nlm.nih.gov/)数据库提供的苹果基因结构注释中，MdMYB88(XP_008389496.1)和MdMYB124(XP_008339925.1)的长度分别为1413bp和1410bp，MdMYB88和MdMYB124基因的核苷酸相似度高达91.49％，氨基酸相似度高达87.95％。
下面结合附图对本发明的应用效果作详细的描述。
MdMYB88/124是苹果中同源基因，功能冗余。利用叶盘转化法，在皇家嘎啦中进行遗传转化，获得转基因苗，MdMYB124过表达株系#1，#2，#19，MdMYB88的过表达株系#23。经过观察两年生的转基因苗和皇家嘎啦苹果比较，发现嘎啦苹果中过表达MdMYB88或者MdMYB124时候，枝条的开张角度比嘎啦苹果树角度大。用量角器进行测量，没有转基因的嘎啦苹果枝条的平均角度为54.25度，MdMYB124过表达株系#1，#2，#19开张角度枝条平均开张角度为79度，74.9度，81.8度，MdMYB88的过表达株系#23平均开张角度为83.6度。沉默株系与嘎啦的枝条夹角没有差异。
并且测量叶片夹角，发现MdMYB88过表达株系叶柄夹角变大。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
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Claims (3)
1.一种增加果树枝条开张角度的转录因子，其特征在于，所述增加果树枝条开张角度的转录因子MdMYB88、MdMYB124；
MdMYB88的核苷酸序列为SEQ ID NO：1；MdMYB88的氨基酸序列为SEQ ID NO：2；
MdMYB124的核苷酸序列为SEQ ID NO：3；MdMYB124的核苷酸序列为SEQ ID NO：4。
2.一种由权利要求1所述增加果树枝条开张角度的转录因子构建的表达载体，其特征在于，所述表达载体：pCAMBIA2300,pGWB414，pGWB415。
3.一种如权利要求1所述转录因子在增加果树枝条开张角度中的应用。
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| 樊松乐 等: "Ｒ2Ｒ3 型 MYB 转录因子 HbMYB88 结构和功能分析", 《植物研究》 * |
| 许瑞瑞等: "苹果基因组中转录因子的鉴定与分析", 《北方园艺》 * |
| 齐思言等: "苹果‘长富2号’开花调控转录因子基因MdIDD7的克隆及表达分析", 《园艺学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114317809A (zh) * | 2022-01-19 | 2022-04-12 | 中国农业大学 | 与苹果树分枝角度相关基因关联的分子标记及其应用 |
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