【作者】 曾亚文；

【导师】 萧凤回；

【作者基本信息】 云南农业大学 ， 作物遗传育种， 2009， 博士

【摘要】 饮食结构不合理导致人类慢性病急剧增加;饮食预防疾病是人类健康的关键。2008年全球糖尿病约2.5亿人,高血脂、高血压和超重均超过10亿人,钙铁锌缺乏影响全球30多亿人口。稻米是全球50%以上人口的主食,也是矿物质和功能食品的重要源泉。稻属核心种质形态、分子评价及其构建技术体系研究相对较多,但核心种质糙米矿质元素和功能成分研究国内外未见报道。云南是中国稻种资源最大的遗传多样性中心,也是优异种质和营养功能稻米资源的富集地区。本研究通过云南稻种初级核心种质的糙米矿质元素含量和进化、功能成分评价及其地带性特征分析,改良品种精米、糙米与土壤元素含量间相关性,并研究了核心种质的表型变异、糙米矿质元素含量及其SSR标记间的关系,具有重要的理论意义和较高实用价值,并取得如下进展:1.首次明确了云南稻种初级核心种质糙米矿质元素含量呈现明显的地带性特征,揭示了籼粳亚种、生态型及其分类性状的系统关系。863份云南稻种初级核心种质糙米P平均含量是土壤总磷的7.17倍,土壤总钾含量是糙米K平均含量的7.56倍。来自云南省5个稻作区16个州市789份地方品种糙米8种矿质元素含量分析显示:糙米元素含量（mg·kg-1）依次为P（3834.83±486.49） > K（2567.72±336.74） > Mg（2567.72±336.74） > Ca（153.67±55.90） > Zn（33.35±13.65） > Fe（32.08±25.51） > Cu（14.22±11.85） > Mn（13.58±3.22）;滇西北糙米P含量高,滇中糙米Ca、Mg、Fe和Zn含量高,滇西南糙米Cu和Mn含量高;糙米高磷钾、中钙镁锰和低铁锌分布区是世界生物多样性最丰富及矿产资源的富集区。总体上789份云南地方稻糙米5种元素（K, Ca, Mg, Fe和Cu）平均含量明显低于94份改良品种,其中Ca元素在籼粳分化增强对冷害的适应性中起了重要作用;地方品种到改良品种随着糙米5种矿质元素的提高而增强了广适性;系统树聚类显示云南稻种分为地方品种和改良品种两类,其中地方品种又分为5个亚组,即A组为紫米,B组为陆稻和光壳稻,C组为糯稻,D组为红米;E组由I组（籼稻、晚稻、白米、地方品种）和II组（粳稻、水稻、非光壳稻、早中稻、粘稻）组成。2.揭示了云南稻种改良品种糙米18种矿质元素含量明显高于精米,且16种元素含量明显低于土壤;微量元素间比大量元素关系密切,其中糙米所含的8种促进人体健康的矿质元素是精米的2倍以上。55个云南省水稻改良品种精米、糙米及其相应的土壤18种矿质元素含量测定显示: 18种矿质元素是功能稻米活性成分的重要部分,其平均含量依次为精米P > K > S > Mg > Ca > Zn > Na > Al > Mn > Fe > Cu > B > Mo > Ni > Sn > Cr > Ba > Sr,糙米P > K > Mg > S > Ca > Zn > Mn > Al > Na > Fe > Cu > B > Mo > Sn > Ni > Cr > Ba > Sr,而土壤则为Fe > Al > Ca > K > Mg > P > S > Mn > B > Na > Ba > Zn > Cr > Cu > Ni > Sn > Mo > Sr;精米和糙米除S、P外16种元素含量均明显低于土壤;发现精米和糙米8种微量元素（Mo、Ni、Cr、Sr、Mn、Zn、Cu、Na）含量间相关性明显比6种大量元素（P、K、Mg、Ca、S和Al）含量间的关系密切。云南土壤以富铁铝高钙为主,而精米和糙米则以磷钾镁硫为主;以精米为主食比糙米更易导致人类健康问题。3.首次发现了云南稻种初级核心种质不同类型间糙米3种功能成分差异显著,呈现明显的地带性特征。996份云南稻种初级核心种质糙米的抗性淀粉、γ-氨基丁酸和总黄酮含量及其栽培型间差异表明:糙米总黄酮含量地方品种极显著高于改良品种,但GABA含量则改良品种显著高于地方品种;抗性淀粉含量则改良品种极显著高于地方品种。其中来自五个稻区16个州市905份地方品种初级核心种质功能成分检测显示:糙米平均抗性淀粉含量（%）为0.75±0.29,滇中极显著高于北部稻作区,滇西北的丽江和滇东北的昭通极显著低于除迪庆州外的13个州市;籼稻显著高于粳稻,粘稻极显著高于糯稻,晚稻极显著高于早中稻,红米和紫米极显著高于白米。糙米γ-氨基丁酸含量(mg·kg^-1)为74.3±25.3,滇南单双季籼稻区明显高于滇西北高寒粳稻区,滇南的思茅、滇中的玉溪和保山至少与5个州市差异显著;水稻极显著高于陆稻,糯稻极显著高于粘稻,晚稻极显著高于早中稻,白米极显著高于红米和紫米。糙米总黄酮含量(mg·kg^-1)为3069.8±1927.5,滇南极显著高于滇中,思茅显著高于8个州市而保山则显著低于7个州市;陆稻极显著高于水稻,粘稻显著高于糯稻,早中稻极显著高于晚稻,红米和紫米极显著高于白米,光壳稻显著高于白壳稻。揭示了糙米总黄酮、γ-氨基丁酸和抗性淀粉含量在水陆、沾糯、早中晚、米色间差异极显著,但籼粳、有无芒和米味间差异不大。4.云南稻种核心种质表型性状、糙米矿质元素含量与其SSR标记有一定相关,首次以SSR标记为基础揭示了糙米矿质元素的含量易受环境的影响,而籽粒的性状主要受遗传基因的控制。基于628份糙米8种元素、23个表型性状及其20个SSR标记间关系研究显示,这些性状组成的620对性状间有182对呈极显著相关;尤其SSR等位基因大小与籽粒性状构成的180对性状中94对呈显著相关,又与穗部性状构成的160对性状中48对呈显著相关。20个SSR标记等位基因大小与一些表型性状呈极显著相关,其中RM224等位基因大小与23个表型性状及8种元素呈显著相关。SSR标记等位基因大小与籽粒或穗部性状间的关系比糙米元素含量和植株性状更密切,籼粳分类的籽粒长宽比及1²节长与14个SSR标记等位基因大小相关显著。以地方稻20个SSR标记的等位基因数目和距离一些级别间糙米矿质元素含量间呈显著差异（P<0.05）;即云南稻种糙米矿质元素含量与基因多样性有关。628份糙米矿质元素含量变异较大,即P为2160⁵500 mg·kg^-1为1130 3830 mg·kg^-1, Ca为61.8⁴88 mg·kg^-1, Mg为864²020 mg·kg^-1, Fe为0.40¹47 mg·kg^-1, Zn为15.1 124 mg·kg^-1, Cu为0.10⁵9.1 mg·kg^-1, Mn为6.7²6.6 mg·kg^-1。因此,检测出云南稻核心种质糙米Ca、Fe和Zn基因型间差异7倍以上,而培育高Ca、Fe和Zn新品种是解决人类矿质营养缺乏的一条经济有效途径。更多还原

【Abstract】 The mankind caused by an irrational diet is dramatic increase in the chronic diseases. The dietary prevention of disease is the key to human health. In 2008, there are approximately 250 million people of diabetes, more than one billion for hyperlipidemia and hypertension as well as overweight people in the world. Ca, Fe and Zn deficiency affect more than 3 billion of the global population. Rice is more than 50% of the staple food, minerals and a source of functional food for the world’s population. Evaluation of phenotype and molecule as well as establishment of technology system on Oryza were relatively more, but elemental elements and functional components in brown rice has not been reported so far. Yunnan is the largest center of genetic diversity as well as rich region of functional and nutrition of rice germplasm resources in China. In this study, their zonal characteristics and evolution of ecotype for the content of mineral elements as well as evaluation of functional components in brown rice of the primary core collection in Yunnan rice, corelation between milled or brown rice and soil elements of improved varieties, the phenotypic variation and mineral elements as well as their inter-relationship between the SSR marker of core collection, which has great theoretical significance and high practical value. The results are as following progress:1.There was fist clear zone characteristics of mineral element contents in brown rice of 863 accessions of primary core collection in Yunnan rice, which revealed that systemic relations including indica-japonica subspecies and ecotype differentiation of mineral elements in classification traits. Average P contents in brown rice of 863 accessions of core collection in Yunnan rice are 7.17 times than that of the total phosphorus in soil, and total potassium content of soil are 7.56 times of average K contents in brown rice. The contents （mg·kg-1） of 8 elements in brown rice of 789 accessions core collection for rice landrace from 16 prefectures of five rice regions in Yunnan Province were in turn P（3834.83±486.49） > K（2567.72±336.74） > Mg（2567.72±336.74） > Ca（153.67±55.90） > Zn（33.35±13.65） > Fe（32.08±25.51） > Cu（14.22±11.85） > Mn （13.58±3.22）. P content in brown rice was the highest from the northwest Yunnan, the highest Ca, Mg, Fe and Zn concentrations in brown rice from the middle Yunnan, and the highest Cu and Mn contents in brown rice from the southwest Yunnan. The distributing zones with the highest （P, K）, middle （Ca, Mg and Mn） and lowest（Fe and Zn） in Yunnan are the enrichment zone of minal resources and most the largest biodiversity. In general, the results show that the mean levels of K, Ca, Mg, Fe and Cu in brown rice for 789 accessions of rice landraces was distinctly lower than that of 94 improved cultivars. They further demonstrate that Ca plays an important role in the differentiation of subspecies indica-japonica, especially to enhance adaptation of cold stress, and that five mineral elements in brown rice enhance the eurytopicity from landrace to improved cultivar. Based on eight mineral elements in brown rice, hierarchical cluster analysis, showed that Yunnan rice could be grouped into rice landrace and improved cultivar, with the rice landrace being further clustered into five subgroups, including group A （purple rice）, group B （upland and nuda）, group C （glutinous）, group D （red rice）, group E divided into two subgroup: I （indica, late, white rice, and landrace） and II （japonica, lowland, non-nuda, early–mid, and non-glutinous）. 2. The contents of 18 mineral elements in brown rice of improve cultivers in Yunnan rice are clear higher than that of milled rice, 16 elements except P and S are clear lower that of corresponding soils. The correlation of microelements in rice are closer than that of macroelements. The first discovered that 8 elements-related health of human being in brown rice is over 2 times than that of milled rice. The contents of 18 mineral elements in milled and brown rice of 55 accessions elite cultivers as well as corresponding soils, were determined by ICP-AES technique. The analytical results showed that 18 mineral elements （S, Mo, Ba, Ni, Fe, Cr, Na, Al, Cu, P, Sn, Zn, B, Mn, Mg, Ca, Sr and K） are the important active compositions of functional rice,their mean content in milled rice are in turn P > K > S > Mg > Ca > Zn > Na > Al > Mn > Fe > Cu > B > Mo > Ni > Sn > Cr > Ba > Sr, brown rice for P > K > Mg > S > Ca > Zn > Mn > Al > Na > Fe > Cu > B > Mo > Sn > Ni > Cr > Ba > Sr, but soil for Fe > Al > Ca > K > Mg > P > S > Mn > B > Na > Ba > Zn > Cr > Cu > Ni > Sn > Mo > Sr; 16 Mineral elements in milled and brown rice （exception S and P） are clear lower than that of soils. The correlation of 8 microelements （Mo, Ni, Cr, Sr, Mn, Zn, Cu and Na） in milled and brown rice are closer than that of 6 macroelements （P, K, Mg, Ca, S, and Al）. There are rich in Fe and Al and Ca in Yunnan soils,but 4 elements （P, K, Mg, and S） give priority to milled and brown rice; The milled rice used for the staple is easier to bring on health problem of human being than that of brown rice.3. The first discovered that there was significantly difference of three functional components in brown rice among different types of primary core collection in Yunnan rice, which showed a clear zone characteristics. The difference of contents and cultivated types of resistant starch andγ-amino butyric acid （GABA） as well as total flavone in brown rice of 996 accessions for primary core collection for Yunnan rice are as follows: Flavonoids contents in brown rice of landraces are the most significantly higher than that of improved cultivars, but GABA content of improved cultivars was significantly higher than that of landraces; resistant starch content of improved cultivars is the most significantly higher than that of landraces. The analytical results showed that the average content （%） for resistant starch in brown rice of 905 accessions from 16 prefectures among five rice regions is 0.75±0.29, it was the most significant highest （p < 0.01） for contents of resistant starch from the middle and South Yunnan province （I, II, III） than that of northern rice regions（V, IV）, and the most significant lowest for contents of resistant starch of Lijiang prefecture of northwestern and Zaotong of northeastern in this province than that of 13 prefectures except Diqing prefectures; Indica is significant higher than japonica, glutinous is the most significant higher than non-glutinous, late is the most significant higher than early–mid, red rice and purple rice is the most significant higher than white rice. The average content （mg?kg-1） for GABA in brown rice is 74.3±25.3, and the content of GABA from South Yunnan single/double cropping rice region is clear higher than that of Northwest Yunnan cold highland japonica rice region; It is significant difference of GABA content in brown rice for that Simao prefecture South Yunnan and Yuxi as well as Baoshan prefecture,at least than that of 5 prefectures; Lowland is the most significant higher than that of upland, non-glutinous is the most significant higher than glutinous, late is the most significant higher than early–mid, white rice is the most significant higher than red rice and purple rice. The average content (mg·kg^-1) for total flavone in brown rice is 3069.8±1927.5, it was the most significant highest （p < 0.01） for contents of total flavone from South Yunnan （II, III） than that of middle Yunnan; It is the most significant highest from Simao prefecture than that of 8 prefectures, but the most significant lowest from Baoshan prefecture than that of 7 prefectures; Upland is the most significant higher than that of lowland, glutinous is significant higher than non-glutinous, early–mid is most significant higher than late, red rice and purple rice are most significant higher than white rice, and nuda was significant higher than non-nuda. These results revealed that most significant difference between lowland versus upland, glutinous versus nonglutinous, early/mid rice versus, and red/purple versus white rice based on the contents of total flavone and resistant starch as well asγ-amino butyric acid （GABA） in brown rice （p < 0.01）, but no significant difference between indica and japonica, awn versus no-awn, common rice and aromatic/soft rice.4. There are some relation between SSR markers and phenotypic traits as well as 8 mineral elements in brown rice of rice landrace in Yunnan Province, which first revealed that mineral elements in brown rice was liable to environmental affect, and grain traits major be controled for genes. It was investigated that allele size of microsatellites associated with phenotypic traits of rice landraces in Yunnan,based on 20 SSR markers and 23 phenotypic traits as well as 8 mineral elements in brown rice within the core collection of 628 accessions; and there was a significant correlation for 182 of 620 pairs among these markers and traits as well as elements. Surprisingly, there was significant correlation for 94 of 180 pairs between allele size of microsatellites and grain traits, and 48 of 160 pairs between allele size of microsatellites and panicle traits. There was a significant correlation between the allele size of 20 SSR markers and some phenotypic traits, such as the significant correlation of 17 pairs between allele size of RM224 and 23 phenotypic traits as well as 8 elements. The allele size of microsatellites was more associated with grain or panicle traits than that of plant traits or element contents in brown rice. Grain length/width ratio and 1―2 internode length, as indica-japonica classification traits, in which two traits were closely associated with the allele size of 14 SSR markers.The 8 elemental concentrations in brown rice among some grades based on number and distance coefficients of alleles for SSR twenty markers for landraces are significantly different （P<0.05）, and further understanding the relationship of mineral elements associated with gene diversity. A large variation in elemental concentrations of brown rice, it was ranged from 2160 to 5500 mg P kg^-1, from 1130 to 3830 mg K kg^-1, from 61.8 to 488 mg Ca kg^-1, from 864 to 2020 mg Mg kg^-1, from 0.40 to 147 mg Fe kg^-1, from 15.1 to 124 mg Zn kg^-1, from 0.10 to 59.1 mg Cu kg^-1, and from 6.7 to 26.6 mg Mn kg^-1. Therefore, genotypic differences of germplasm evaluations for Ca, Fe, and Zn concentrations in rice grain had be detected up to over seven times, which suggesting that selection for high Ca, Fe and Zn cultivars was a effective approach.更多还原