转多基因水稻植株的获得

将2-3个外源目的基因与筛选标记基因hpt连在同一个载体上,用基因枪法对1个粳稻品种和4个籼稻品种进行了转化,粳稻中花8号转化频率最高为56.4%,灿稻七丝软占、特青、奇妙香、九六零转化频率分别为6.4%,2.5%,2.8%和1.7%,分子杂交实验证明,通过两次潮霉素抗性筛选的T0代再生植株100%含有hpt基因,其中70%以上含有外源目的基因的完整表达结构。遗传分析表明,大部分T1代植株中潮霉素抗敏比符合盂德尔3：1的分离规律。     

基因枪法转基因水稻中hpt基因稳定遗传

基因枪转化将潮霉素磷酸转移酶基因（ｈｐｔ）导入粳稻品种７７１７０,获得可育的转基因植株,研究外源基因遗传的稳定性。自交后代（Ｔ１和Ｔ２）经潮霉素筛选获得抗性植株和敏感植株,分子鉴定结果表明抗性植株带有ｈｐｔ基因,而敏感植株中没有ｈｐｔ基因存在。Ｔ１和Ｔ２代中潮霉素抗性表现为显性单基因位点的遗传方式,符合孟德尔分离规律,并得到分子鉴定结果的证实。Ｓｏｕｔｈｅｒｎ杂交结果显示,ｈｐｔ基因多拷贝整合在水     

农杆菌介导寒地水稻遗传转化的研究

通过农杆菌介导法,将拟南芥转录激活因子ICE1基因导入寒地品种垦鉴稻10号成熟胚愈伤组织中,将经潮霉素筛选得到的抗性愈伤进行分化,获得了一些再生抗性植株,部分植株经PCR检测为阳性,证明外源目的基因已整合到水稻的染色体上,从而建立了高效水稻遗传转化体系。实验表明:YEB和MS按适当比例混合作为悬浮培养基转化率较高;干燥除菌效果和转化率均优于羧苄青霉素、头孢霉素液体除菌;潮霉素采用低压-高压-低压方式筛选最好;AS和葡萄糖均能提高转化率。     

籼稻基因枪转化的研究

对国内外１７个籼稻品种进行了基因枪转化,研究了有利于籼稻愈伤组织诱导和生长的培养条件和筛选程序,１５个品种获得了潮霉素抗性愈伤组织,５个品种再生了植株,包括当前国际上推广的ＩＲ系统及国内的优良品种和恢复系,分子生物学证据证明潮霉素基因已经整合入籼稻的基因组。最高植株转化频率接近一般粳稻水平,多数低于粳稻水平。这为建立籼稻的转化系统打下了基础。     

通过根癌农杆菌介导的共转化获得具有蜡质基因反义片段的转基因水稻

在构建剔除潮霉素抗性基因的反义蜡质基因重组质粒p13W8的基础上,用分别携带p13W8质粒和有潮霉素抗性基因的pCAMBIA1300质粒的根癌农杆菌,在不同处理条件下对粳稻3个品种进行共转化.试验结果显示,共感染混合液中pCAMBIA1300菌液比例高,抗性愈伤组织的转化频率也较高;抗性愈伤组织的PCR检测结果表明,潮霉素抗性基因的阳性检测率为98%,反义蜡质基因的阳性检测率为68%.在105株转基因植株中,反义蜡质基因阳性株为30株,占转基因植株的28.6%.乙酰丁香酮处理组的平均转化频率略高于未处理组;在乙酰丁香酮的各种处理中,直接浸泡愈伤组织的转化频率稍高于其它处理方式.     

根癌农杆菌介导的水稻转化及转基因R1代植株表型特征

用根癌农杆菌介导的方法对２ 个粳稻品种和４ 个籼稻品种进行了转化。粳稻成熟胚和籼稻幼胚来源的愈伤组织用携带质粒ｐＧＩＨ 的农杆菌ＥＨＡ１０１ 感染,对所有品种均获得较高的愈伤转化频率（２０．８３ ％ ～６２ ．３２ ％ ） 。粳稻“申香粳４ 号”植株转化频率为１７ ．３９ ％ ,“秋丰”为９ ．２１％ 。４ 个品种籼稻中仅“超丰早１ 号”获得１ 株转化植株。Ｓｏｕｔｈｅｒｎ 杂交分析、ＧＵＳ 染色、ＴＤＮＡ 整合边界序列分析等结果表明外源基因整合入植物基因组中。对Ｒ１ 代分析结果表明外源基因能遗传给后代,大多数株系的分离比符合３∶１ ,也有少数株系分离比异常。对转基因Ｒ１ 代水稻植株表型特征分析结果显示转化植株株高下降,每株分蘖数减少,每株籽粒数明显减少,部分植株播种至开花所需时间延迟。     

农杆菌介导类胡萝卜素合成酶基因LycB转化水稻的研究

以3个水稻品种的成熟胚诱导的良好胚性愈伤组织为受体,以LycB为目的基因,应用根癌农杆菌介导法对水稻进行遗传转化,同时以抗性愈伤率为依据,对影响转化的几个因素进行优化研究。结果表明：预培养4d、侵染5～10min、农杆菌菌液浓度OD600值0.7～1.0、共培养2d有利于提高转化率。经潮霉素筛选获得的抗性植株经PCR和PCR-Southern分析鉴定,初步证明外源基因LycB已整合到水稻的基因组中。     

提高水稻基因枪转化效率的研究

在转化前将受体材料转移到高渗培养基上,即用含有一定浓度的甘露醇和山梨醇的培养基对受体材料进行处理。基因枪轰击后把受体材料转出,结果表明,这样做明显提高了瞬时表达（ＧＵＳ基因的表达）效果,用可筛选的标记基因（潮霉素磷酸转移酶基因,ＨＰＴ）试验表明,经高渗处理后,稳定转化的效率也有提高。用基因枪将带有ＨＴＰ基因的粒子轰击其盾片,经过筛选及植株再生,获得了转基因植株,ＰＣＲ扩增反应及Ｓｏｕｔｈｅｒｎ杂交证明了外源基因已整合入水稻的基因组     

hpt与bar基因作为水稻转基因筛选标记的比较研究

标记基因的选择是影响植物遗传转化和转基因后代筛选成败的关键因素。hpt与bar作为两种常用的水稻转化筛选标记被广泛应用于水稻的转化。为比较两者在实际应用中的效果, 文章首先对比了在潮霉素和除草剂(Bialaphos)两种筛选剂下水稻遗传转化的情况。研究表明, hpt基因的转化筛选体系相对于bar基因在转化效率上提高近两倍, 转化周期提前至少10 d, 且插入基因拷贝数更低。随后, 文章分析了利用潮霉素浸种法在田间筛选转基因水稻的可行性, 研究显示当潮霉素浓度大于167 mg/L时, 可以对以水稻品种kitaake为亲本的转基因材料进行有效筛选, 达到常规除草剂的筛选效果。但与除草剂相比较, 潮霉素的田间筛选成本却处于劣势。文章研究和讨论了hpt和bar基因在遗传转化和后代田间筛选中的优缺点, 并提供了一种利用潮霉素浸种法筛选转基因后代阳性植株的手段, 为将来在水稻转基因研究工作中根据实际需求选择合适的遗传转化、筛选体系提供参考。     

水稻几丁质酶基因导入芥菜型油菜的研究

以芥菜型油菜 (BrassicajunceaL .)下胚轴为转化材料 ,通过根癌农杆菌 (Agrobacteriumtumefaciens)介导将水稻的几丁质酶基因 (Ricechitinasegene)导入“泸洲四陵”油菜品种中 ,获得抗潮霉素的再生转基因植株 ,并讨论了影响油菜再生及转化效率的几个因素。对部分经潮霉素筛选得到的再生植株进行了多次重复PCR检测 ,发现其中 40 %以上的潮霉素抗性植株均表现出较强的阳性反应 ,初步证明几丁质酶基因已整合到油菜细胞核基因组中。     

新型四倍体水稻创制及其杂种优势利用研究进展

新型四倍体水稻是本研究团队创制的一类高育性多倍体水稻新种质,携带有多种优异基因。该类材料与同源四倍体水稻杂交获得的杂种F₁产量优势明显,具有应用前景。本文首先概述了同源四倍体水稻的研究进展,并重点阐述了新型四倍体水稻创制过程、主要特点、杂种优势表现及其涉及的分子遗传机理,最后提出未来研究的思路及需要解决的问题,为加快新型四倍体水稻在多倍体水稻育种中的利用提供依据。     

籼、粳杂种的“随体丢失”

1985—1986,观察籼、粳及籼,粳F_1杂种终变期核仁染色体数,籼为2个二价核仁染色体,粳为1个二价核仁染色体,籼、粳F_1杂种核仁染色体数与父本水稻的核仁染色体数相同。由此,讨论了“随体丢失”在水稻育种,粳稻起源及遗传学方面的意义。     

Critical Yield Factors for Achieving High Grain Yield in Early-Season Rice Grown under Mechanical Transplanting Conditions

Double-season rice cropping systems occupy a large portion of the rice production area in southern China. Because the problem of insufficient labor, mechanical transplanting (in contrast to the manual transplanting) was become more attractive in double-season rice system. However, the decisive yield factors which resulting in high grain yield of early-season rice are unclear under mechanical-transplanted conditions. In present study, the field experiments were conducted in the early season in 2017 and repeated in 2018 in Santang Town, Hunan Province, China. Ten early season rice cultivars (Zhuliangyou 819, Lingliangyou 268, Lingliangyou 104, Luliangyou 996, Xiangzaoxian 24, Xiangzaoxian 32, Xiangzaoxian 45, Xiangzaoxian 42, Zhongjiazao 17, and Zhongzao 39) were used as materials in this study. The difference in grain yield and closelyrelated agronomic and physiological traits of ten tested cultivars were compared. The range of yields (t ha^–1 at 86% dry matter) in 2017 was 6.2 to 8.7 (mean 7.8) and in 2018 was 6.5 to 8.4 (mean 7.8). Grain weight and pre-heading biomass accumulation had potent significant positive correlations with the grain yield. The greater pre-heading biomass accumulation was major attributed to higher apparent radiation use efficiency. Our results suggested that early-season rice cultivars to achieve the high grain yield in mechanical-transplanted conditions depends on apparent radiation use efficiency in the pre-heading period and higher grain weight.     

Arsenic tolerance, uptake and translocation by seedlings of three rice cultivars

By simulating the anaerobic conditions with agar nutrient solutions, effect of arsenic (As) on the growth and As uptake by hybrid, conventional and glutinous rice cultivars were studied. It showed insignificant effect of As on the root dry weights of three rice cultivars when treated by As of different concentrations. The shoot dry weights of hybrid and glutinous decreased with As concentrations increasing, while low concentrations of As (0.5 mg L^?1) could enhance the growth of conventional rice. Generally, As concentrations in roots and shoots increased as As concentrations of treatment solutions increasing. The root system had strong ability to uptake and accumulate As. The root As concentrations ranged from 156 to 504 mg kg^?1, representing 63.40%–81.90% of the total As concentrations in rice, which were much higher than shoot As concentrations. The fact that the glutinous rice had higher biomass, higher tolerance, and lower As concentrations in its roots and shoots than the other two rice cultivars proved that the glutinous rice was more applicable to As-polluted soils.     

不同水稻品系幼苗对砷（As）的耐性、吸收及转运

利用琼脂培养基模拟水稻生长的厌氧环境,研究了As对不同水稻品系幼苗生长的影响以及As在其体内的积累及转运特性.结果表明,不同浓度(0～4.0mg/L)的As对供试水稻品系根部干物质积累无显著影响(P>0.05).杂交稻与糯稻的地上部干物质积累随基质中As浓度的增加呈减小趋势,但低剂量的As(0.5mg/L)促进常规稻的生长.水稻地上部的As积累量随基质中As浓度的升高总体均呈增加趋势.水稻根系对As具有较强的吸收与累积能力.水稻根部As的积累量为156.31～504 03mg/kg,占总As含量的63.40～81.90%,远远高于其地上部As的积累量.相比于其它两个品系,糯稻的生物量积累高,耐性指数较大,根部及地上部对As的积累量较低,因此更适合种植在As污染土壤.     

籼稻品种浙辐802抗瘟性遗传研究

通过与８个日本鉴别品种杂交和利用日本代表菌系研５４－０４及我国南方稻区菌系９５-ｈ接种鉴定,研究了籼稻品种浙辐８０２的抗性遗传。研究结果表明,该品种对菌系研５４-０４的抗性由 ２对显性基因控制,对菌系 ９５-ｔ２的抗性由 １对显性基因控制。基因等位性分析确认,浙辐８０２中抵抗９５－ｔ２的抗病基因与Ｐｉ－ｉ基因等位,与Ｐｉ－ａ、Ｐｉ－ｓｈ、Ｐｉ－ｋ、Ｐｉ－ｚ、Ｐｉ－ｂ、Ｐｉ－ｔ、Ｐｉ－ｔａ等已知基因位点为非等位关系;抵抗菌系研５４－０４,感染菌系９５-ｔ２的基因与Ｐｉ－ｉ、Ｐｉ－ｋ、Ｐｉ－ｂ、 Ｐｉ－ｔ ４个已知基因位点为非等位关系。对这个基因与其他已知基因的等位性进行了分析,认为它可能是１个未被命名的新基因。     

Cadmium,copper, and zinc levels in the rice and rice field soil of Houston,Texas

Fifty-one pairs of hulled rice samples and the soil from which each rice sample was grown were analyzed for heavy metals in August, 1979, in order to estimate the background contamination of cadmium (Cd), copper (Cu), and zinc (Zn) in rice grown in the Houston, Texas area. Both samples were divided into three groups by soil types and colors. The cadmium concentration in Texas rice was only one-half to one-quarter lower than that of Asian rice. However, the levels of Cu and Zn in rice in Texas were similar to those reported. Soil heavy metals were lower than ever reported, but these values were consistent with the geochemical characteristics of the Texas Houston area. No particular relationship was found between the three metals in rice and the metals in soil where the sampled rice was grown.     

Origin,dispersal, cultivation and variation of rice

There are two cultivated and twenty-one wild species of genus Oryza. O. sativa, the Asian cultivated rice is grown all over the world. The African cultivated rice, O. glaberrima is grown on a small scale in West Africa. The genus Oryza probably originated about 130 million years ago in Gondwanaland and different species got distributed into different continents with the breakup of Gondwanaland. The cultivated species originated from a common ancestor with AA genome. Perennial and annual ancestors of O. sativa are O. rufipogon and O. nivara and those of O. glaberrima are O. longistaminata/, O. breviligulata and O. glaberrima probably domesticated in Niger river delta. Varieties of O. sativa are classified into six groups on the basis of genetic affinity. Widely known indica rices correspond to group I and japonicas to group VI. The so called javanica rices also belong to group VI and are designated as tropical japonicas in contrast to temperate japonicas grown in temperate climate. Indica and japonica rices had a polyphyletic origin. Indicas were probably domesticated in the foothills of Himalayas in Eastern India and japonicas somewhere in South China. The indica rices dispersed throughout the tropics and subtropics from India. The japonica rices moved northward from South China and became the temperate ecotype. They also moved southward to Southeast Asia and from there to West Africa and Brazil and became tropical ecotype. Rice is now grown between 55°N and 36°S latitudes. It is grown under diverse growing conditions such as irrigated, rainfed lowland, rainfed upland and floodprone ecosystems. Human selection and adaptation to diverse environments has resulted in numerous cultivars. It is estimated that about 120000 varieties of rice exist in the world. After the establishment of International Rice Research Institute in 1960, rice varietal improvement was intensified and high yielding varieties were developed. These varieties are now planted to 70% of world's riceland. Rice production doubled between 1966 and 1990 due to large scale adoption of these improved varieties. Rice production must increase by 60% by 2025 to feed the additional rice consumers. New tools of molecular and cellular biology such as anther culture, molecular marker aided selection and genetic engineering will play increasing role in rice improvement.     

不同浓度Al3+胁迫对水稻根生长的影响

铝毒害是酸性土壤限制农作物产量的主要因素之一。本文以水稻为材料,探讨了不同浓度的AlCl3(0、50、100、150 mmol.L-1)胁迫处理对水稻幼苗根生长的影响。结果表明,随着Al3+浓度和胁迫时间的增加,水稻幼苗根的生长受到抑制,长度和鲜重都减小,根尖细胞死亡的程度增大,根中H2O2含量上升;与对照(未经DMTU预处理)相比,经dimethylthiourea(内源过氧化氢抑制剂)预处理的水稻幼苗在Al3+胁迫下根细胞死亡程度减小,H2O2含量降低,根的生长抑制得到缓解,表明内源H2O2的积累是造成Al3+对水稻幼苗根生长抑制的原因之一。