The Source of Genes Related to Rice Grain Starch Synthesis Among Cultivated Varieties and Its Contribution to Quality

The property of starch in rice grain endosperm is a very important determinant for rice quality, and it is essential to understand the genetic effect of the genes related to starch synthesis in high-yielding rice varieties for rice quality improvement. The physicochemical properties （e.g., amylose content, gel consistency, and RVA profile） were assessed on 53 rice varieties, including certain typical indica/japonica landraces and certain high-yielding modern varieties. And molecular markers for Sbel, Sbe3 developed on the basis of sequence diversities between the rice subspecies indica and japonica, together with PCR-Acc I marker for Wx gene were used to investigate the genotypes of 53 rice cultivars. The result showed that the developed molecular markers for Wx, Sbel, Sbe3 could distinguish indica or japonica alleles at three loci. Among all the 53 rice cultivars, six genotypes were observed when Sbel, Sbe3, and Wx loci were considered together, while the genotypes of WxiSbelJSbe3i and WxiSbelJSbe3J were absent. In order to explore the genetic effects of the three genes, especially for starch branching enzyme genes, ANOVA and multiple comparison analysis were conducted. The results showed that rice cultivars with different genotypes exhibited different phenotypes, including amylose content, gel consistency and certain RVA characteristics, and the significant differences among the six genotypes were observed. It was concluded that these three genes had randomly recombined during the process of the rice variety development. And the genetic effects of indica and japonica alleles at three gene loci were different, of which, Wx gene plays a major role in determining the starch properties, followed by Sbel and Sbe3, and the genetic effects of Sbel and Sbe3 in different backgrounds （Wx~, WxJ） are different. The results have provided a clue for rice good quality variety development, and the molecular markers will benefit to the improvement in quality of rice.

Breeding of Selectable Marker-Free Transgenic Rice Lines Containing AP1 Gene with Enhanced Disease Resistance

In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene （HPT） were cloned into the two separate T-DNA regions of the binary vector pSB130, respectively, and introduced into the calli derived from the immature seeds of two elite japonica rice varieties, Guangling Xiangjing and Wuxiangjing 9, mediated by Agrobacterium-mediated transformation. Many cotransgenic rice lines containing both the AP1 gene and the marker gene were regenerated and the integration of both transgenes in the transgenic rice plants was confirmed by either PCR or Southern blotting technique. Several selectable marker-free transgenic rice plants were subsequently obtained from the progeny of the cotransformants, and confirmed by both PCR and Southern blotting analysis. These transgenic rice lines were tested in the field and their resistance to disease was carefully investigated, the results showed that after inoculation the resistance to either bacterial blight or sheath blight of the selected transgenic lines was improved when compared with those of wild type.

QTL Mapping for Rice RVA Properties Using High-Throughput Re-sequenced Chromosome Segment Substitution Lines

The rapid visco analyser （RVA） profile is an important factor for evaluation of the cooking and eating quality of rice. To improve rice quality, the identification of new quantitative trait loci （QTLs） for RVA profiling is of great significance. We used a japonica rice cultivar Nipponbare as the recipient and indica rice 9311 as the donor to develop a population containing 38 chromosome segment substitution lines （CSSLs） genotyped by a high-throughput re-sequencing strategy. In this study, the population and the parent lines, which contained similar apparent amylose contents, were used to map the QTLs of RVA properties including peak paste viscosity （PKV）, hot paste viscosity （HPV）, cool paste viscosity （CPV）, breakdown viscosity （BKV）, setback viscosity （SBV）, consistency viscosity （CSV）, peak time （PET） and pasting temperature （PAT）. QTL analysis was carried out using one-way analysis of variance and Dunnett＇s test, and stable QTLs were identified over two years and under two environments. We identified 10 stable QTLs： qPKV2-1, qSBV2-1; qPKV5-1, qHPV5-1, qCPV5-1; qPKV7-1, qHPV7-1, qCPV7-1, qSBV7-1; and qPKV8-1 on chromosomes 2, 5, 7 and 8, respectively, with contributions ranging from -95.6% to 47.1%. Besides, there was pleiotropy in the QTLs on chromosomes 2, 5 and 7.

Transgenic Expression of the Recombinant Phytase in Rice (Oryza sativa)

In most of the cereal crop, phytic acid is the main storage form of phosphorus, which can decrease the bioavailability of phosphate. Transgenic expression of phytase is regarded as an efficient way to release phosphate from phytate in transgenic plants. In this study, a plant expression vector, containing the recombinant phytase gene driven by the maize ubiquitin （Ubi） promoter was constructed and introduced into an elite rice variety via Agrobacterium-mediated transformation. During the experiment, a total of 15 independent transgenic rice lines were regenerated. The results of PCR and Southern blot indicated that the target gene was integrated into the genome of transgenic rice plants. Moreover, the RT-PCR analysis of total RNAs extracted from the immature seeds of several transgenic lines showed that the recombinant phytase gene could be normally expressed. The inorganic phosphorus content, both in the mature seeds and the leaf was significantly higher in the transgenic plants than in the untransformed wild type.

RL3(t),Responsible for Leaf Shape Formation,Delimited to a 46-kb DNA Fragment in Rice

Two mutants with rolled leaves, temporally designated as rl3（t）-I and rl3（t）-2, were served for exploring the mechanism underlying the rolled leaf characteristic. Except for having typical rolled leaves, the plant heights and panicle lengths of rl3（t）-1 and rl3（t）-2 significantly decreased, and the seed-setting rate also decreased when compared with wild type 93-11. Cytological analysis suggested that the rolled leaf phenotype might be caused by the changes of number and size of bulliform cells. Genetic analysis indicated rl3（t）-1 is allelic to rl3（t）-2, and controlled by a recessive gene. Gene mapping result indicated that RL3（t） gene resided in a 46-kb long region governed by the sequence tag site markers S3-39 and S3-36 on rice chromosome 3. The result provides an important clue for further cloning the RL3（t） and understanding the mechanism of rice leaf development.

PDCA循环联合营养支持对有机磷中毒患者心理睡眠质量及生命质量的影响研究

目的探究戴明循环(PDCA循环)联合营养支持对有机磷中毒患者的影响。方法对湖州市中医院2017年2月~2018年2月收治的80例有机磷中毒患者开展对照研究,按照随机数字表法分为观察组(PDCA循环联合营养支持)与对照组(PDCA循环),每组40例,探讨实施对不同护理效果的影响。结果护理前研究对象在ADL评分、ALB上差异无统计学意义(P>0.05),观察组护理第1周、第2周后在ADL评分上明显比对照组低(P<0.05);观察组护理第1周、第2周后ALB水平明显比对照组高(P<0.05)。研究对象护理前的睡眠质量、睡眠效率、入睡时间、睡眠药物、睡眠时间、日间功能、睡眠障碍评分比较差异无统计学意义(P>0.05),两组护理后的睡眠质量、睡眠效率、入睡时间、睡眠药物、睡眠时间、日间功能、睡眠障碍评分均低于同组护理前(P<0.05)。观察组护理后的睡眠质量、睡眠效率、入睡时间、睡眠药物、睡眠时间、日间功能、睡眠障碍评分低于对照组(P<0.05)。护理前研究对象的HAMA、HAMD评分差异无统计学意义(P>0.05),观察组在护理后的在HAMA、HAMD评分上明显比对照组低(P<0.05)。在护理满意度上两组相比,观察组(97.50%)明显比对照组(85.00%)高(P<0.05)。结论将PDCA循环联合营养支持应用于有机磷中毒患者后,可明显改善患者负性心理,提升生活质量,改善睡眠质量及营养状态。

宫颈癌患者血清相关指标与临床病理特征及预后的相关性分析

目的探讨宫颈癌患者血清相关指标与临床病理特征及预后的相关性。方法选取2015年1月—2019年1月在浙江中医药大学附属湖州中医院接受治疗的116例宫颈癌患者为观察组;另选取进行健康体检的42例健康者为对照组。使用双抗夹心酶联免疫分析法检测两组研究对象血清中基质金属蛋白酶9(MMP-9)的含量;使用全自动化学发光免疫分析仪测定血清T细胞免疫球蛋白黏蛋白分子4(Tim-4)、程序性死亡配体1(PD-L1)水平;统计观察组患者年龄、病理类型、病例分级、临床分期、肿瘤直径及肿瘤浸润深度和淋巴结转移情况。采用多因素和Cox对因素回归性分洗宫颈癌患者预后的相关性。结果观察组患者血清中MMP-9、Tim-4及PD-L1水平明显高于对照组(P<0.05)。观察组患者血清中MMP-9表达与病理分级、浸润深度及淋巴结转移相关(P<0.05),观察组患者血清中Tim-4、PD-L1水平的表达与TNM分期、病理分级、浸润深度及淋巴结转移相关(P<0.05)。通过单因素分析结果显示TNM分期、病理分级、浸润深度、淋巴结转移、血清中MMP-9、Tim-4及PD-L1水平与宫颈癌患者预后有关(P<0.05)。COX回归性分析发现TNM分期、病理分级、浸润深度、淋巴结转移、血清中MMP-9、Tim-4、PD-L1水平均为宫颈癌患者预后的独立危险因素(P<0.05)。结论宫颈癌患者血清中MMP-9、Tim-4、PDL1水平、TNM分期、病理分级、浸润深度及淋巴结转移的阳性表达均为影响预后的危险因素。