色氨酰-tRNA合成酶基因WRS1调控水稻种子发育

【目的】氨酰-tRNA合成酶（aminoacyl-tRNA synthetases, aaRSs）与遗传信息传递密切相关,已发现植物中aaRSs家族蛋白在维持翻译功能之余,还参与配子发生与胚发育、质体的早期发育以及免疫信号的感知与病害防御等生物学过程。本研究利用水稻胚乳发育缺陷突变体,分析水稻色氨酰-tRNA合成酶（WRS1）在胚乳发育中的作用,证明WRS1基因编码一个影响水稻胚乳发育的关键因子。【方法】本研究通过甲烷磺酸乙酯（ethyl methane sulfonate, EMS）诱变籼稻（Oryza sativa subsp. indica）品种N22,筛选到一个稳定遗传的水稻粉质胚乳突变体（wrs1）,图位克隆获得目标基因。对wrs1成熟种子进行形态学观察以及淀粉相关理化性质测定,利用细胞学切片分析wrs1发育中胚乳的结构,利用实时荧光定量PCR（quantitative real-time PCR, qRT-PCR）和GUS活性染色分析基因表达模式,通过qRT-PCR比较野生型与突变体花后12 d胚乳中淀粉合成相关基因表达情况,免疫印迹检测野生型与突变体成熟种子中淀粉合成酶蛋白积累情况,使用全自动氨基酸分析仪测定游离氨基酸含量。【结果】 wrs1突变体幼苗表现出明显的发育滞后且逐渐蔫萎死亡,从杂合突变体（WRS1wrs1）中分离到的粉质籽粒呈现明显的腹部皱缩,粒厚、千粒重下降,同时总淀粉含量下降,糊化淀粉的峰值黏度和崩解值均低于野生型。wrs1突变体发育胚乳中复合淀粉颗粒变小,排列疏松。WRS1定位于第12染色体长臂约183 kb的区间内,测序发现编码色氨酰-tRNA合成酶（tryptophanyl-tRNA synthetase, WRS）基因的第6外显子上发生单碱基替换,导致一个保守位置上的甲硫氨酸被替换。wrs1突变体中大部分淀粉合成相关基因表达量下调,且野生型与突变体间基因表达的变化与相应蛋白积累的差异存在不一致的趋势。wrs1突变体籽粒中蛋白质积累降低,而游离氨基酸含量显著升高。【结论】 WRS1编码色氨酰-tRNA合成酶,该基因突变后通过影响氨基酸稳态和蛋白质合成,造成淀粉合成相关基因异常表达从而影响淀粉的合成与积累,导致种子发育缺陷。     

Strategies for applying selenium for biofortification of rice in tropical soils and their effect on element accumulation and distribution in grains

This study aimed to investigate the effects of the selenium (Se) application via soil (SeVI) and leaf (SeIV and SeVI) on agronomic attributes of rice and the spatial distribution of Se and other elements in biofortified rice grains. For this, a field trial was conducted using six Se rates (0, 5, 10, 20, 40, and 80 g ha⁻¹). Physiological evaluations in rice leaves, rice grain yield, and Se and other elements content were assessed. Spatial distributions of Se and other elements were visualized mapping the biofortified rice grains with μXRF. The results showed that the strategies and rates of Se applied to the plants were effective for producing Se-rich rice grains. Although Se application did not change the rice yield, it affected some enzyme activities, improving the antioxidant system of the plant. The spatial distribution of the different elements found in the biofortified rice grains varied substantially, with Se accumulations mainly in the endosperm, whereas P, K, Fe, and Zn accumulate in the embryo. Agronomic biofortification of rice with Se, via soil or leaf, is a promising strategy to be adopted in tropical soils in order to produce rice grains with adequate Se contents for human consumption.     

松树种子胚乳DNA的抽提与分析

运用改良SDS(十二烷基硫酸钠)法,从马尾松、黄山松、黑松等松树种子胚乳中提取到较高纯度的DNA,并对其纯度、浓度及产率进行了分析.经RAPD检测表明,DNA扩增效果良好,完全能满足常规DNA实验的要求,为针叶树种等其它小粒种子DNA的提取提供了经济、快速、可靠的实验方法.     

水稻胚乳突变体筛选与特性研究进展

本文在简要介绍胚乳的发育、组成及品质性状遗传的基础上,综述了与淀粉、蛋白质和微营养相关的系列胚乳突变及相关性状的基因定位。胚乳突变主要包括糯性、暗色、云雾状、糖质、皱缩、粉质米、低直链淀粉、高直链淀粉、低粘滞性、高抗性淀粉、低水溶性蛋白、蛋白体缺失、高赖氨酸、高铁、低植酸等方面。     

优质蛋白玉米选育的分子遗传途径

本文综述了opaque突变基因及其修饰基因的最新研究进展。从分子生物学角度分析了优质蛋白玉米育种过程中突变基因及修饰基因改善蛋白质品质和控制籽粒性状的遗传机理,提出了利用突变基因及修饰基因减少玉米醇溶蛋白,增进玉米品质的思路。     

Relationships among the chalkiness,kernel size and endosperm cell morphology of rice kernels at different spikelet positions within a panicle

The occurrence of chalky kernels in rice is causally related to kernel size and endosperm morphology. This study aimed to investigate the occurrence of chalky kernels by analyzing kernel size, and the number and area of endosperm cells. Spikelets were sampled from upper and middle primary branches and lower secondary branches in a panicle, and divided into four categories: upper, middle, and lower perfect (PF) kernels and lower milky-white (MW) kernels. On the lower secondary branches, there was a higher percentage of chalky kernels, with smaller kernel lengths, widths and thicknesses, than the kernels on the upper and middle primary branches. MW kernels were smaller in size than PF kernels even on the same lower secondary branches. Regardless of grain appearance quality traits, the total areas of endosperm cross sections in lower kernels were significantly smaller than in upper kernels owing to the decreased cell area, and there was a significant negative correlation between the number of cells and average cell area. When the numbers and the areas of cells were analyzed using angular 30° intervals from the line connecting the center point and the dorsal vascular bundle, the MW kernels had significantly less cells than PF kernels near the ventral side at 120–180°. Thus, the decrease in the number of cells near the ventral side was a main causal factor in the decrease in MW kernel widths compared with PF kernel widths, and this suggested that cell division in MW kernels was inhibited at the early grain-filling stage.     

Unravelling transcriptome changes between two distinct maize inbred lines using RNA-seq

Seed size play a significant role in maize yield production. Two maize inbred lines with distinct seed size and weight, V671 (a large-seed inbred line) and Mc (a small-seed inbred line), were investigated by RNA-seq at 14 days after pollination (DAP), when maize endosperm undergoes an active transition from mitosis to storage accumulation. RNA-seq expression data showed that the small-seed line Mc had a higher storage accumulation activity, whereas the large-seed kernel line V671 possessed a higher DNA synthesis activity. An investigation of the pattern of increasing kernel width at serial DAPs showed that V671 experienced an increased kernel width later than did Mc, but the rate and duration of increase were longer in V671. SDS-PAGE of the storage proteins and quantitative RT-PCR of cell cycle-related genes and indole-3-acetic (IAA) synthesis genes certified that the transition from mitosis to storage accumulation starts earlier in Mc. We hypothesized that the difference in the mitosis-to-storage accumulation transition accounts for the larger seed size in V671 vs. Mc.     

Effects of Shading at Different Stages After Anthesis on Maize Grain Weight and Quality at Cytology Level

The objective of the current study is to investigate the effects of different stages of shading after anthesis on grain weight and quality of maize at cytology level.The shading experiments were conducted in the field from 2005 to 2006,with a common maize cultivar(TY2)as the experimental material.Plants were given stress using horizontal shading net and the light intensity was reduced by 55%.Field-grown maize plants were shaded at 1-14 d(S1),15-28 d(S2),and 29-42 d(S3)after pollination,respectively.Control plants(S0)were grown under natural light.Grain weight,quality,endosperm cell proliferation,cob sugar content,and grain pedicel vascular bundle cross section area were measured.The ultrastructural changes of endosperm cells and endosperm transfer cells were observed after pollination.The result indicated that the grain weight,starch content,endosperm cell number,and volume were declined after shading.On the contrary,the proportion of embryo and endosperm,protein content,and fat content in grain increased.Shading treatments significantly delayed the development of the starch granules and remarkably reduced the endosperm filling status.Among the three treatments.the number of the grain endosperm was the least under shading stress at 1-14 d after pollination.However,the volume of starch granules and the substantiation of endosperm under shading treatment at 15-28 d after pollination were the worst.Compared with the control(natural sunlight without shading),the soluble sugar of maize cob increased significantly,while there was no obvious change in vascular structure of small cluster stalk.The number of protein body in maize endosperm was influenced markedly by low light at different stages after pollination.Low light decreased the volume of the grain endosperm transfer and the cell wall extensions of the basal transfer cells became thinner and shorter under shading treatment than those of the control.Furthermore,the degree of connection and the capacity of the nutrient transport were decreased and the mitochondrion number of the transfer cell was reduced after shading.The change in grain quality after shading was observed due to increase in the proportion of embryo and endosperm.The morphology and functions in endosperm transfer cell and the shortage of energy restricted the nutrient transport greatly with shading at different stages,suggesting that an impeded flux may be one of the important reasons for the reduction of maize grain weight of maize grain at later growth stage under low light condition.     

四个不同粒重水稻品种颖果发育的比较

 以粒重差异较大的4个水稻品种为供试材料,采用树脂切片、酶解胚乳细胞和显微观察等方法,比较研究了品种间在颖果生长、胚乳细胞增殖、果皮和胚乳结构等方面的差异,探讨了影响颖果生长的因素。 大粒品种颖果发育时间较小粒品种长,其胚乳细胞数、胚乳干质量及单个胚乳细胞平均干质量均高于小粒品种。在粒重相近的情况下,籼稻颖果发育和淀粉积累快于粳稻。与小粒品种相比,大粒品种子房壁细胞中淀粉粒多,子房壁细胞生长的持续时间长,果皮及背部维管束衰亡迟。 小粒品种胚乳外层细胞在花后7 d已转化成糊粉层细胞,大粒品种胚乳外层细胞要在花后10 d才转化成糊粉层细胞。 大粒品种的库容大和生理活性期长是其颖果能显著增大的生理原因。     

基于透射光和反射光图像同位分割的小麦品种识别方法研究

为提高基于机器视觉的小麦品种识别准确性,本文通过透射光和反射光同位图像分割对种子颜色特征参数进行了优化提取.采用透射光图像辅助反射光图像分割的方式从种子图像中分割出胚部区域,并分别提取小麦整粒、种胚、胚乳区域的颜色特征参数.以济麦22、济麦44、京麦9、京麦11共4个品种种子作为研究对象,利用HALCON机器视觉软件获...