Pea-Fer基因导入与表达对水稻植株和子粒重要矿质元素积累的影响

为研究外源豌豆铁蛋白基因(Pea-Fer)的导入和表达对水稻植株和子粒重要矿质元素的积累所产生的影响,本试验以原始受体亲本粳稻品种秀水11为轮回亲本,外源豌豆铁蛋白转基因纯系Fer34为非轮回亲本,采取连续回交,结合GUS标记基因辅助选择技术,在BC6F3获得含Pea-Fer的Fer34-XS11,它与秀水11构成一对近等基因系.并利用该近等基因系,研究Pea-Fer基因导入对水稻植株不同生长发育阶段(幼苗期、分蘖期、成熟期)的根、茎(叶鞘)、叶及子粒不同部分(谷壳、米糠、糙米、精米)的主要矿质元素(Fe、Ca、Mn、Zn)积累的影响.结果表明:外源豌豆铁蛋白基因(Pea - Fer)的导入,使得水稻植株在不同生长发育时期的根、茎(叶鞘)、叶等器官中的Fe含量较对照品种秀水11明显增加,而对于Ca、Mn和Zn的含量并没有显著影响;同时,水稻子粒中Fe含量积累也有较高提升,而Ca、Mn和Zn的积累却未出现显著变化.这为深入开展转基因富铁水稻新种质的研究和利用提供了一定依据.     

外源钙（Ca）对毛葱耐镉（Cd）胁迫能力的影响

以对环境敏感的毛葱(Allium cepa var.agrogarum L.)为材料,通过水培试验分别研究不同浓度Ca(0、0.1、1、10 mmol/L)对Cd(10μmol/L、100μmol/L和300μmol/L)胁迫下毛葱幼苗生长、叶片光合特性、体内Cd积累和矿质营养的变化,探讨Ca缓解敏感植物Cd毒害的生理生化机制。结果表明:(1)Cd显著抑制了毛葱的生长并导致其根端弯曲、发黄,叶片绿色加深;外源Ca显著削弱了Cd的毒害,缓解了其对毛葱生长的抑制;(2)Cd导致毛葱叶片光合色素含量大幅度上升却显著降低了毛葱的光合作用;外源Ca延缓了光合色素上升的速度,提高了毛葱的净光合速率、气孔导度和蒸腾速率;(3)Cd胁迫导致毛葱体内Cd含量显著增加,并造成明显的矿质营养失衡,主要表现为显著降低了毛葱根中Mg、Mn,叶中Ca、Mg、Mn、Zn等元素的含量,毛葱根中Ca、Fe、Zn,叶中Fe元素含量的显著增加,扰乱了毛葱体内矿质营养的内稳态;外源Ca削弱了毛葱对Cd的积累,减轻了Cd胁迫所造成的矿质营养失衡。因此,外源Ca能通过抑制Cd的吸收,促进叶片光合作用及气体交换速率,维持植物体的含水量、植物叶片光合色素含量及矿质营养的平衡等途径来增强毛葱对Cd胁迫的耐性。     

苦参凝集素蛋白基因转化水稻的研究

以水稻杂交品种‘云资粳41号’为受体材料,通过农杆菌介导法将苦参凝集素蛋白基因(SFL)导入水稻细胞,采用氯酚红法和PCR检测外源基因是否整合到水稻基因组中。结果显示:外源基因成功转入水稻基因组,并获得一批转基因水稻植株;转基因植株叶片离体接种稻瘟病菌的检测结果显示,转基因植株与对照(非转基因植株)相比有明显的抗性,证明SFL基因在水稻中得到表达。研究表明,基于SFL基因所具备的广谱抗菌作用,可以预期所得转基因水稻植株很可能对水稻的多种病原菌具有良好的抗性,为选育新的抗稻瘟病水稻新品种以及拓宽栽培稻抗病遗传基础增加抗稻瘟病基因奠定了基础。     

毛果苔草湿地营养元素的积累、分配及其生物循环特征

毛果苔草(Carex lasiocarpa)湿地地上部分积累量小于地下部分的积累量,在地上几个构件中,叶片比叶鞘积累量大,而穗的积累量最小;地下部分中细根比根茎的积累量大.土壤分室营养元素贮量在系统的各分室中占绝对优势,毛果苔草湿地土壤中各种营养元素总贮量的顺序为K＞Fe＞N＞Ca＞P＞Mg＞Mn＞Zn＞Cu;它们的吸收系数的排序是Mn＞N＞P＞Zn＞Mg＞Cu＞Ca＞Fe＞K;几种营养元素的利用系数的排序是Mn＞N＞P＞Zn＞Mg＞Cu＞Ca＞Fe＞K;几种营养元素的循环系数的排序为Ca＞K＞Mg＞N＞P＞Mn＞Zn＞Cu＞Fe;因此,该系统中钙、钾的存留比例最小,而流动性较大,而铁则相反,存留比例大,流动性较小.     

水稻籽粒矿质元素含量遗传及主要农艺性状相关性分析

通过了解水稻子粒钙、镁、铜、铁、锌和硒等矿质元素含量,以绿旱1号作为父本与102S、KD36S、7HS012及7HS013不育系为母本进行配组,对父本与杂交后代主要农艺及经济性状、水稻子粒中的矿质元素含量进行了遗传分析,并对各杂交后代的主要农艺性状和子粒中矿质元素含量以及各矿质元素含量间进行了相关性分析.结果表明,杂交后代农艺及经济性状多数表现出超高亲分离现象,且LK3的农艺及经济性状均表现为最好.水稻子粒矿质元素含量与部分农艺和经济性状存在明显的正相关或负相关关系,且K、Mg、Zn、Se等元素含量与水稻产量密切相关.水稻子粒中矿质元素含量大小依次为P＞K ＞Mg ＞S＞ Ca＞ Mn ＞Fe ＞Zn ＞Cu ＞Se,4个杂交后代的K元素含量表现出超高亲分离现象,LK2、LK3和LK4的Fe元素含量也表现出超高亲分离现象.水稻子粒大量与微量元素之间相关性最高,微量元素之间相关性次之,大量元素之间相关性最小,以7HS012不育系为母本得到的杂交后代农艺性状表现较好,矿质元素含量最高,7HS013次之,其他不育系表现一般.本研究为选育出耐旱及营养价值高的水稻新品种母本的选择提供了理论依据.     

生物炭对水稻苗期生长、养分吸收及土壤矿质元素含量的影响

为明确生物炭对东北粳稻苗期的生长发育和对育苗基质营养含量的影响、探明生物炭在东北冷凉地区水稻育苗生产上的应用潜力,以东北稻田土壤为基础、生物炭为外源添加物(5.0%~20.0%,w/w),研究生物炭对东北冷凉区水稻秧苗株高、干物质积累量、矿质元素N、C、S、P、K、Mn、Zn、Cu、Fe、Mg、Ca和Na的吸收及育苗基质中矿物质元素含量变化的影响。结果表明:在育苗基质中添加生物炭对水稻苗期株高、干物质积累有显著影响;其中施用5.0%、10.0%和15.0%生物炭时株高分别比对照平均提高1.2%、5.0%和6.0%,干物质积累量分别比对照平均提高18.2%、27.7%和43.4%;水稻秧苗随着生物炭施用量增加,N、C、S、P、K、Mn、Ca、Na、Mg、Zn、Cu和Fe吸收量均呈先升后降趋势;在施炭量为5.0%时矿物质元素Mn、Zn和Cu出现峰值,在施炭量为10.0%时矿物质元素Fe出现峰值,在施炭量为15.0%时矿物质元素N、C、S、P、K、Ca、Na和Mg出现峰值;在育苗基质中,N、C、S、P、Zn、Cu、Mg和Na含量随生物炭用量增加均呈上升趋势,而K和Fe随生物炭量增加,其含量逐渐减少,Mn和Ca含量无明显规律性变化;在育苗基质中施用10.0%~15.0%的生物炭能够促进水稻秧苗生长,有利于水稻秧苗对大部分矿物质元素的吸收与利用,同时提高育苗基质固定与保持一些重要营养元素的能力。     

农杆菌转化获得转B．t．基因水稻及其生物学鉴定

以水稻主栽品种"秀水11"和"春江11"预培养4d未成熟胚为转化受体,经农杆茵LBA4404／pGBI4A2B(含B．t．基因)感染后,筛选出抗性愈伤组织并获得转化植株。其中抗性愈伤组织产生频率达44％～70％,转化植株产生频率达27％～64％。转基因植物总DNA经PCR和Southernblot分子杂交试验表明,T-DNA上的外源基因已整合到水稻的基因组里。对转B．t．基因水稻植株进行了两种水稻主要害虫纵卷叶螟和二化螟的饲虫试验。与对照相比,7d后纵卷叶螟幼虫死亡率达31％～49％,同时纵卷叶螟对转基因水稻叶片的危害程度大大减轻。二化螟在咬食转B．t．基因水稻植株7d后的校正死亡率达15％～71％,30d后多数转基因水稻仍能正常抽穗,表明转基因水稻对上述害虫具有一定抗性。     

豌豆凝集素和血红蛋白基因对水稻的转化和表达

为了扩大根瘤菌的突破产范围和试探根瘤菌在非豆科植物上的固所为作用,将豌豆凝集素基因（pl）和Parasponia andersonii血红蛋白基因 （phb）构建在同一个植物表达载体上,用基因枪法将其导入水稻（Oryza sativa L.ssp.japonica）。经PCR扩增和Southern杂匀分析,证明外源目的基因已整合到水稻基因组中。GUS组织化学染色及豌豆凝集素基因的Western印迹实验和表达产物的原位杂交,证实外源基因在转基因水稻中表达。在40个转化植株中18株有pl和phb基因的PCR产物,得率为45％。再用18株植物做pl基因的Western blot检测,有3株有翻译表达,占40株的7.5％,18株的17％。为水稻与根瘤菌的相互作用和固氮作用的可能性研究奠定了一定的基础。     

红花对土壤矿质元素的吸收和积累

采用电感耦合等离子体原子发射光谱法(ICP-AES)检测红花不同部位和时期以及栽培土壤中K、Ca、Na、Mg、Fe、Cu、Zn、Mn、Cr、Cd、Pb、Ba、Ni和Al元素的含量。结果表明:红花各部位矿质元素含量差异明显。在红花各不同部位中K、Fe、Zn元素在幼苗中含量最高,Ca、Mg元素在成熟叶中含量最高,Cu元素在种子中含量最高;而K和Ca元素在种子中含量最低,Mg、Fe、Cu和Zn元素分别在花、茎、成熟叶和茎中含量最低。红花的幼苗和花对K元素吸收积累明显,幼苗和种子对Cu元素吸收积累明显,Ca元素在成熟叶中积累较多,相应元素含量远比土壤中相应的元素含量高。红花对土壤不同矿质元素的吸收利用差异较大,在长期栽培红花的土壤中应注意对吸收利用多的元素结合施肥进行补充。     

IaaL基因的维管束特异表达导致烟草茎和根外植体分化能力的改变

我们利用水稻苯丙氨酸解氨酶启动子(AQ630)和吲哚乙酰胺赖氨酸酯合成酶基因(iaaL)构建了维管束特异表达的植物表达载体pBAL1并导入烟草基因组中。比较了对照植株、转基因植株的幼茎和根外植体在组织培养中分化能力的变化,结果表明转基因植株茎外植体的不定芽形成明显受到促进,而转化植株根外植体在不定根发生方面对外源IAA的敏感性下降。     

Analysis of Metal Element Distributions in Rice (Oryza sativa L.) Seeds and Relocation during Germination Based on X-Ray Fluorescence Imaging of Zn,Fe, K,Ca, and Mn

Knowledge of mineral localization within rice grains is important for understanding the role of different elements in seed development, as well as for facilitating biofortification of seed micronutrients in order to enhance seeds’ values in human diets. In this study, the concentrations of minerals in whole rice grains, hulls, brown rice, bran and polished rice were quantified by inductively coupled plasma mass spectroscopy. The in vivo mineral distribution patterns in rice grains and shifts in those distribution patterns during progressive stages of germination were analyzed by synchrotron X-ray microfluorescence. The results showed that half of the total Zn, two thirds of the total Fe, and most of the total K, Ca and Mn were removed by the milling process if the hull and bran were thoroughly polished. Concentrations of all elements were high in the embryo regions even though the local distributions within the embryo varied between elements. Mobilization of the minerals from specific seed locations during germination was also element-specific. High mobilization of K and Ca from grains to growing roots and leaf primordia was observed; the flux of Zn to these expanding tissues was somewhat less than that of K and Ca; the mobilization of Mn or Fe was relatively low, at least during the first few days of germination.     

Genetic Identification of Quantitative Trait Loci for Contents of Mineral Nutrients in Rice Grain

In present study, Fe, Zn, Mn, Cu, Ca, Mg, P and K contents of 85 Introgression linee (ILs) derived from a cross between an elite indica cultivar Teqing and the wild rice (Oryza rufipogon) were measured by inductively coupled argon plasma (ICAP) spectrometry. Substantial variation was observed for all traits and most of the mineral elements were significantly positive correlated or independent except for Fe with Cu. A total of 31 putative quantitative trait loci (QTLs) were detected for these eight mineral elements by single point analysis. Wild rice (O. rufipogon) contributed favorable alleles for most of the QTLs (26 QTLs), and chromosomes 1, 9 and 12 exhibited 14 QTLs (45%) for these traits. One major effect of QTL for zinc content accounted for the largest proportion of phenotypic variation (11%-19%) was detected near the simple sequence repeats marker RM152 on chromosome 8. The co-locations of QTLs for some mineral elements observed in this mapping population suggested the relationship was at a molecular level among these traits and could be helpful for simultaneous improvement of these traits in rice grain by marker assisted selection.     

籼型黑米粒形性状的遗传效应及其与矿质元素含量的遗传相关性

采用禾谷类作物种子数量性状的遗传模型,分析了灿型黑米稻品种双列杂交F1和F2种子的粒重,粒长,粒宽和粒长／粒宽等粒形性状的遗传效应及其与米粒中矿质元素Fe,Zn,Mn和P含量的遗传相关性,结果表明：4种粒形性状同时受制于种子直接遗传效应,母体效应和细胞质作用影响,其中种子直接遗传效应比母体效应和细胞质效应的作用更大,且种子直接遗传效应以加性效应占主导,粒重,粒宽和粒长／粒宽的种子直接遗传率较高,杂种早代单粒选择效果较好,粒长的种子直接遗传率和母体遗传率均属中等,较高世代的杂种进行单株选择和单粒选择均有一定效果,4种粒形性状与其米粒中矿质元素Fe,Zn,Mn和P含量表现较强的种子直接加性相关,直接显性相关,细胞质相关,母体加性相关和母体显性相关,在特种稻育种实践中,可以通过粒形性的间接选择,达到改良其矿质元素含量等营养品质性状的目标。     

Effect of exchangeable sodium percentage and presubmergence on yield and nutrition of rice under field conditions

Summary The effects of exchangeable sodium percentage (ESP) levels of 82, 72, 65 and 35 and 0, 15 and 30 days of presubmergence (submergence prior to the transplanting of rice) on yield and chemical composition of rice and availability of Fe, Mn, Zn and P in soil were studied factorially in a field experiment. Presubmergence increased rice yields at all ESP levels, the effect being more pronounced at high ESP's. Increasing ESP decreased yields and the contents of Ca, Mg, K, Fe, Mn, Zn and Cu but increased that of P and Na in the crop. Presubmergence enhanced absorption of all the above elements by the crop except P, K, Mg, Zn and Cu in the grain and decreased Na in grain and straw. Growing of rice under submerged conditions also facilitated the improvement of these soils. Effects of submergence and ESP on the availability of Fe, Mn, Zn and P in soil and their role in the nutrition of rice are discussed. The results suggest that 15 to 30 days presubmergence improved rice yields on a calcareous sodic soil of the Indo-Gangetic alluvial plain.     

Contribution of mineral nutrients from source to sink organs in rice under different nitrogen fertilization

The pot experiment with three treatments of nitrogen (N) topdressing was performed with the japonica rice cultivar viz. Huaidao 5. Remobilization of nine mineral nutrients including N, phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), manganese (Mn), and copper (Cu) was measured from the source organs including bracts, leaf, and sheath to sink rice grain. Experimental results showed considerable contribution of bracts to grain for N, Mg, and Zn, with the averages contributions of 5.96, 12.56, and 12.34%, respectively, indicating a positive role of rice bracts in N, Mg, and Zn remobilization during grain filling. By contrast, minor contribution of bracts to grain P, K, and Cu was revealed, with the contribution rate being 0.99, 3.90, and 3.05%, respectively. Further, a net increase in Ca and Fe concentrations of bracts was detected, implying that bracts function as a sink of these mineral nutrients. In addition, grains produced at a moderate level of N topdressing had higher Fe and similar Zn concentration in comparison with those at high N level, suggesting the possibility of N management for maintaining Fe and Zn level under high yielding conditions.     

Iron accumulation does not parallel the high expression level of ferritin in transgenic rice seeds

To answer the question whether iron accumulation in transgenic rice seeds depends on the expression level of exogenous soybean ferritin, we generated two kinds of ferritin hyper-expressing rice lines by introducing soybean ferritin SoyferH-1 gene under the control of the rice seed storage glutelin gene promoter, GluB-1 and the rice seed storage globulin gene promoter, Glb-1, (GluB-1/SoyferH-1 and Glb-1/SoyferH-1, DF lines), and by introducing the SoyferH-1 gene under the control of Glb-1 promoter alone (Glb-1/SoyferH-1, OF lines). Ferritin expression was restricted to the endosperm in both lines and protein levels determined by western blot analysis were up to 13-fold higher than in a construct previously reported FK22 (GluB-1/SoyferH-1, in genetically Kitaake bachground); however, the maximum iron concentrations in seeds of both of the new lines were only about 30% higher than FK22. The maximum iron concentration in the OF and DF lines was about threefold higher than in the non-transformant. The mean Fe concentration in leaves of ferritin over-expressing lines decreased to less than half of the non-transformant while that the plant biomasses and seed yields of the ferritin-transformed lines were not significantly different from those of the non-transformant, suggesting that accumulation of Fe in seeds of hyper-expression ferritin rice did not always depend on the expression level of exogenous ferritin but may have been limited by Fe uptake and transport. No obvious differences were observed for other divalent-metal concentrations (Ca, Cd, Cu, Mg, Mn and Zn) in the seeds among all experimental lines and non-transformant.     

太阳辐射减弱对水稻光合生理特性和中微量元素积累的影响

近年来雾霾等大气环境污染问题突出,气溶胶导致到达地表的太阳辐射强度降低,对农作物生长造成了较大影响。为了探讨太阳辐射强度降低对粮食作物生理特性和矿质元素营养的影响,本研究以‘南粳5055’水稻品种为供试材料,采用田间随机区组设计,通过不同程度遮光处理模拟弱光环境(CK,不遮光对照;Y₁、Y₂遮光率分别为19%、45%),研究了在水稻关键生育期(拔节期、抽穗期、灌浆期)内叶绿素含量(SPAD)、叶面积指数(LAI)、叶片净光合速率、产量和籽粒中、微量金属元素(Ca、Mg、Fe、Zn、Mn、Cu)含量的响应特征。结果表明: 全生育期内遮光处理抑制了水稻光合作用产物的合成,降低了叶面积指数;但在生育前期对叶绿素含量无显著影响,在生育后期则显著增加。两种遮光处理下水稻的千粒重较CK分别降低了14.4%和18.4%,结实率降低了4.3%和12.9%,从而导致水稻产量下降,且随着遮光程度增加,水稻减产率加大,产量分别下降了58.5%和66.4%。遮光处理籽粒糙米和颖壳中金属元素含量升高,尤其微量元素含量显著上升。可见,遮光对水稻生长产生了不利影响,最终使其减产,并使Cu、Mn等重金属元素含量增加,这可能造成污染风险进而对人体健康造成威胁,因此,太阳辐射减弱对作物产量和品质的影响需要综合评价。     

密穗型和散穗型粳稻品种品质特性比较

比较2个粳稻密穗型和散穗型品种(系)品质结果表明,穗型间和穗型内品质存在较大差异,2个密穗型品种(系)平均垩白特性不及2个散穗型品种.密穗型新品系EA6垩白粒率较相同穗型品种秀水63低约1/2,与散穗型品种秀水11相仿,较散穗型品种越光高约1倍;垩白度较秀水63和秀水11低约1/2,较越光高1倍多.调查穗部不同部位籽粒垩白表明,穗基部籽粒垩白对品种总垩白影响最大,而这种影响密穗型更大于散穗型.4个品种饭香味相仿,光泽、软度、粘度、食味和冷饭软度等指标EA6显著或极显著优于秀水11,秀水63不及秀水11;EA6多数指标接近越光.表明密穗型品种经改良其品质有可能达到甚至超过散穗型品种.     

黑米中矿质元素铁、锌、锰、磷含量的遗传效应研究

采用禾谷类作物种子胚乳数量性状模型,分析黑米稻品种双列杂交Ｆ１和Ｆ２种子的Ｆｅ、Ｚｎ、Ｍｎ、Ｐ含量的遗传效应。结果表明,４种矿质元素含量同时受制于种子直接遗传效应、母体效应和细胞质作用影响。其中,Ｆｅ、Ｚｎ、Ｍｎ含量的种子直接效应比母体效应和细胞质效应的作用更大;Ｐ含量则主要受种子直接加性、母体加性和显性效应共同作用。Ｆｅ、Ｚｎ、Ｍｎ含量的种子直接遗传率较高,在杂种早代分别结合农艺性状选择单株上各