【作者】 姚立新；

【导师】 康向阳； 续九如；

【作者基本信息】 北京林业大学 ， 林木遗传育种， 2010， 博士

【摘要】 冬枣(Zizyphus jujuba Mill. cv. Dongzao)是我国特有的优良晚熟、鲜食枣品种。在冬枣产业迅速发展的过程中,出现不同产地争当“第一”、盲目引种、果实品质下降等问题。本研究针对以上问题,在山东沾化、河北黄骅和沧县三地设立产地对比试验园,将选自不同产地(沾化、黄骅、庆云、沧县和乐陵)的冬枣在同一立地条件下栽培,对冬枣果实和果核表型、果实营养组成、感官评价、抗寒性和光合生理指标等进行了连续4年的测定分析,探讨了冬枣产生差异及品质下降的真正原因；本文还进行了冬枣cDNA文库构建、测序和生物信息学分析；研究了冬枣主要病虫害的无公害防治措施并探索了冬枣杂交育种技术。主要研究成果如下：1.多点、多年观测结果表明,在同一年份、同一试验园内,不同产地冬枣在果实和果核表型、果实营养组成和感官评价等方面没有明显差异；而在不同年份、不同试验园之间,冬枣果实表型和品质各项指标差异显著。说明影响冬枣果实表型和品质的主要因素是环境条件和栽培管理措施,而不是产地来源。就冬枣群体而言,在长期栽培过程中,由于芽变和人工选择,仍存在着个别单株的差异。所以,冬枣改良的方向不是产地选择,而是单株选择。2.冬枣叶片N／P变化范围为11.11～13.54,属于N含量制约的植物类型。冬枣叶片N、P、K元素含量在7月～9月初比较稳定,这段时间是上述3种元素最佳营养诊断采样期。3.冬枣的光合性状受环境的影响变化较大,测定结果与测定时间、地点甚至部位等均有很大关系。沧州地区不同产地冬枣叶片Pn的日变化呈较为明显的不对称双峰曲线,年变化呈单峰曲线,冬枣为中午光合速率降低型植物。4.温度的骤变是可能引起冬枣冻害的一个重要原因,应当根据各地实际情况注意防护；冬枣在适生区栽植也要防止个别年份出现极端低温时遭受冻害；温度条件不适合的地区不可盲目引种,进行栽培时应采取保护措施。5个不同产地冬枣抗寒力有一定差别,但在不同指标间表现不一致。不同产地冬枣在沧州地区均能正常生长,无冻害发生。5.利用3种不同发育时期的冬枣果实构建混合cDNA文库,从该cDNA文库中随机挑取单克隆进行5’端单向测序,得到1060条EST序列,经过cross-match和CAP3软件处理后共获得628条Unigene。在Uniprot数据库中比较发现有534条Unigene与已知基因具有同源序列,占Unigene总数的85%。利用ESTPiper在线分析软件对Unigene进行GO功能基因分类,发现273条可以在“分子功能”、“生物学过程”、“细胞内组分”得到分类。鉴定出MYB转录因子、肉桂醇脱氢酶、甘露糖转运等与果实品质形成相关功能基因以及ACC氧化酶、超氧化物歧化酶、热激蛋白等与冬枣果实后熟相关功能基因。利用PERL程序对628条Unigene进行EST-SSR分析,结果表明在116条EST序列中包含有147个SSR,以二核苷酸和三核苷酸为主要重复类型。6.在调查已有栽培技术和自己试验的基础上,本研究总结课题组多年工作,结合生产实践,制定了《绿色食品(AA级)冬枣栽培技术规程(草案)》,为实施冬枣无公害栽培提供了依据和工作细则。7.冬枣人工杂交育种困难,本研究中杂交试验失败的主要原因是冬枣坐果率低、种仁率低、胚败育,而且杂交时授粉时期晚、部分品种花期不遇,并遭遇绿盲蝽象危害等,应开拓枣树育种的新思路。更多还原

【Abstract】 Dongzao jujube(Zizyphus jujuba Mill. cv. Dongzao) is regarded as the best late maturing and fresh cultivars of Chinese jujubes. With the rapid development of Dongzao jujube’s industry in recent years, such problems as the degradation of fruit quality, trying to be first, blind introduction happened. Three comparison orchards, which located at Zhanhua county in Shandong province, Huanghua and Cangxian county in Hebei province were established in order to probe the cause of fruit degradation and variation of quality. Such indexes as the fruit quality, the fruit stone phenotype, components of fruit nutrients, food sensory evaluation, cold hardiness and photosynthetic physiology were analyzed at these three comparison orchards for 4 years.The paper also carried a jujube cDNA library construction, sequencing and bioinformatics analysis; pollution-free prevention and the feasibility of cross breeding of Dongzao jujube were also studied in this work. Following was the key results in this thesis.1. There was no qualitative difference for Dongzao jujubes of different provenances in the same comparison orchard and in the same year. But the difference of Dongzao jujube between different orchards and different years was significant. This results showed that the phenotypic and fruit quality of Dongzao jujube were mainly contributed by environmental conditions, cultivation and management measures, but not by provenances. Because the long-term effect of mutant and artificial selection, there are still variations for individual plant. The direction of genetic improvement was individual selection not group selection.2. The N/P of the leaf of Dongzao jujube ranges from 11.11 to 13.54, which indicated Dongzao jujube was a kind of vegetation type influenced by the content of N. The contents of N, P, K in Dongzao jujube leaf were stable from July to September, which was the optimal sampling period for nutrition diagnostic.3. Photosynthetic characteristics of Dongzao jujube were greatly influenced by the environment varation, which were different with the sampling time, place and sampling part. The diurnal variation of Pn in Cangzhou region showed obvious asymmetry double curve, while annual change showed a single peak curve, Dongzao jujube was decline photosynthetic rates plant at noon.4. The sudden change of air temperature is an important factor which leads to freezing of Dongzao jujube, so the protection should be based on the actual situation in different areas. Freezing damage which was caused by extreme low temperature even should be to prevented in Dongzao jujube normal region. Blind introduction should be avoided, especially for those places which temperature conditions are not suitable for the cultivation of Dongzao jujube. Some protective measures were necessary for cultivation. There was some difference between different provenances of Dongzao jujube, but no trees suffered from freezing injury under natural situation in Cang County, Hebei Proviance of China.5. The fruit of three different development stages were equally mixed to construct a fruit cDNA library.1060 EST sequences were obtained by one run sequencing from 5’ end which was randomly picked from cDNA library.628 Unigenes were obtained after analyzed by cross-matching and CAP3 software. Compared with Uniprot database,534 Unigene sequences were identified with known homologous genes, accounting for 85% of the total number of Unigene. A GO analysis was carried out by ESTPiper, an online software, which resulted that 273 Unigene sequences could be classified in the "molecular function", "biological process" and "cellular component". Some fruit quality related genes, such as MYB transcription factor, Cinnamyl alcohol dehydrogenase, GDP-mannose transporter, and some genes associated with jujube fruit ripening genes, such as the ACC oxidase, superoxide dismutase, heat shock protein, also could be identified. Furthermore, the Unigene sequences were used for EST-SSR analysis, which revealed 147 SSRs in 116 EST sequences. The dinucleotide and trinucleotide repeats were found as the main motif types.6. Based on pollution-free pest control and pollution-free cultivation, "Technical specification for the fruit production of Ziziphus jujube cv. Dongzao at green food AA grade " was compiled, which provides a theoretical basis and working instruction for the pollution-free cultivation of Dongzao jujube.7. There were difficulties in artificial crossbreeding for Dongzao jujube. The main reasons for failure of crossbreeding were asynchronous flowering of the related cultivars, delaying pollination, low fruit-setting rate, very low kernel percentage, embryo abortion, and damage induced by Lygus bug and so on.更多还原