Effect of Rolled Leaf Gene Rl_(t) on Grain Quality in Hybrid Rice

Effects of rolled leaf gene Rl（t） on grain quality characters of hybrid rice were analyzed by using three pairs of rolled leaf near-isogenic lines under two fertilizer treatments. Under normal fertilizer level （e.g. 450 kg urea per ha）, head rice rates and milled rice recovery of rolled leaf hybrids were significantly higher than those of corresponding non-rolled crosses, while the chalky rice rate and chalkiness were all lower. Of the RVA profiles, the peak viscosity, the hot paste viscosity and the breakdown viscosity of the rolled were all higher than those of the corresponding non-rolled ones to various degrees. Increasing fertilizer application for promoting panicle development increased the brown, milled and head rice rates except for Shanyou 63, furthermore, significant difference of head rice rates existed between the rolled leaf Shanyou 559 and Shanyou 559; while the peak viscosity, the hot paste viscosity and the breakdown viscosity all decreased to different levels; changes of values of other characters had no apparent regularity. It suggested that Rl（t） could improve rice quality under certain conditions.

Analysis of Gene Effect on Four Characters of Immature Embryo Culture in Maize

This study was to find the regularity in the hereditary variation for the main culturing characters of the immature embryo culture in maize. Two kinds of inbred-line, R18-599 （red） with very excellent embryo culturing capacity and R15 with very poor embryo culturing capacity, were used as P1 and P2 for obtaining six generations. By culturing immature embryos of the six generations, four culturing characters, namely embryonic callus induction efficiency, nonembryonic callus induction efficiency, cloning ability of the embryonic callus, and number of regenerating plants, were analyzed using the general mean analysis and generation joint analysis. Results showed that the embryonic callus induction efficiency accorded with two major additive-dominance-epistatic genes and polygene-mixed additive-dominance-epistatic inheritance model. The induction efficiency of the nonembryonic callus accorded with two major additive-dominance-epistatic genes. The number of regenerating plants accorded with one major gene and polygene-mixed additive-dominance inheritance model. The cloning ability of the embryo callus accorded with two major genes and polygene-mixed inheritance model, whereas the effect of epistatic gene on this character was identified results of the two methods, generation joint analysis may genetic information. to be different using the two methods. By comparison of the not only raise experimental precision but also provide more

Effect of apolipoprotein E gene Hha Ⅰ restricting fragment length polymorphism on serum lipids in cholecystolithiasis

AIM To investigate the role of apolipoprotein E (apoE) polymorphism in the lithogenesis of gallstone and the hereditary pathogenesis of the disease.METHODS Polymerase chain reaction (PCR)was used to study apoE phenotypes and allelefrequencies in patients with gallstones and control, and the fasting serum lipids of subjectswere also measured by enzymatic methods.RESULTS The levels of triglyceride (TG) andvery low density lipoprotein cholesterol (VLDLC) were much higher in Ez/, patients than that inE,/, control. E,/, patients were accompanied withremarkably low levels of high density lipoproteincholesterol (HDLC) and its subforms. But in E,/#patients there were only slight changes in levelsof VLDLC and low density lipoprotein cholesterol (LDL--C).CONCLUSION Different apoE phenotype patientswith gallstones have different cheracteristics ofdyslipidemia and the average level of serum lipids in patients with gallstones are higher thansubjects without gallstones in the same apoEgene phenotype. EZ allele is possibly one of thedangerous factors in the lithogenesis of cholecystolithiasis.

GENETIC MODELS AND ANALYSIS METHODS FOR SEX-LINKED AND MATERNAL GENE EFFECTS

Genetic models are proposed for analyzing sex-linked and maternal effects as well as autosomal gene effects.For the model with no genotype×environment interaction,the total genetic effect is partitioned into direct additive （A）,direct dominance （D）,sexlinked （L）,maternal additive （Am） and maternal dominance （Dm） genetic components.For the model including genotype×environment interaction （GE）,GE can also be partitioned into components of direct additive by environment interaction （AE）,direct dominance by environment interaction （DE）,sex-linked by environment interaction （LE）,maternal additive by environment interaction （AmE ）,and maternal dominance by environment interaction （DmE）.Linear functions of genetic components are listed for parent,F1,and F2.A set of parents,their reciprocal F1’s and F2’s is applicable for efficient analysis.Variance and covariance components can be well mated by MINQUE（O/l） with the jackknife procedure.The t-test conducted by the jackknife procedure is applicable for detecting significance of variation.Adjusted Unbiased Prediction （AUP） method is suggested for predicting genetic effects.

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.

Isolation, Identification and Earliness Effect Analysis of Rice Dominant Earliness Gene Ef-cd

Ef-cd gene is a dominant earliness gene located on the short arm of rice chromosome 3. In this paper, through continuous backcross, self-pollination and molecular marker assisted selection, individual Ef-cd gene was isolated and its nearly isogenic lines were constructed by using early-maturing indica line 6442S-7 as the donor parent, and by using latematuring indica line Minghui 63 （MH63）, Shuhui 881 （SH881） and Shuhui 527 （SH527） as the recurrent parents （genetic background）, respectively. Further, it was found out that Ef-cd gene could generally advance rice to head 11-14 d earlier. So, it was considered that Ef-cd gene played an important role in rapid developing early-maturing and super high-yielding rice varieties.

Analyses of gene effects and inheritance of resistance to bacterial blight in rice

We analyzed the resistant inheritance of ajaponica variety,Jia23,to two bacterial blightpathogen strains,KS-6-6 and Zhe 173,rep-resenting respectively the two predominantpathogenic types(Ⅱ and Ⅳ)in the rice crop-ping area along the Yangtze River Valley.Jia23 was crossed with susceptible vari-eties,Ewan8 and 7416.Fplants were back-

Determining Nodulation Regulatory (Rj) Genes of Myanmar Soybean Cultivars and Their Symbiotic Effectiveness with <i>Bradyrhizobium japonicum</i>USDA110

Soybean (Glycine max L.) plays an essential role in human nutrition as a protein source, and in plant nutrition as a N source. The rate of N fixation varies depending on the cultivars and compatibility between the inoculated Rhizobium strain and the host cultivar. Characterizing the nodulation regulatory (Rj) genes is necessary to determine the compatibility of cultivars and Rhizobium strains. Rj genes were previously identified based on inoculation tests and PCR analyses. The six cultivars Yezin-3, Yezin-7, Yezin-11, Shan Seine (Local), Madaya (Local), and Hinthada (Local) were identified as harboring the Rj4 gene. Two cultivars, Yezin-6 and Yezin-8, were classified as non-Rj-gene harboring. Two other cultivars, Yezin-9 and Yezin-10, were identified as Rj3- and Rj2Rj3-gene harboring, respectively. Ours is the first report on Rj3- and Rj2Rj3-gene harboring cultivars in Myanmar. We evaluated Myanmar soybean cultivars for symbiotic effectiveness, relying on the standard strain Bradyrhizobium japonicum USDA110. In our first experiment, the soybean cultivar Yezin-11 (Rj4) showed the highest N fixing potential. Based on their potential for fixing N and nodulation, the top six soybean cultivars were Yezin-11 (Rj4), Yezin-9 (Rj3), Yezin-6 (non-Rj), Yezin-8 (non-Rj), Yezin-3 (Rj4) and Yezin-10 (Rj2Rj3). These cultivars were selected for a second experiment, which revealed that the N fixation, nodulation, and plant growth of Yezin-11 (Rj4) *Corresponding author. A. Z. Htwe et al. 2800 were superior to the other cultivars. We conclude that Yezin-11 (Rj4) is the most efficient cultivar for nodulation and N fixation when inoculated with B. japonicum USDA110.

Hitchhiking Effect Mapping: A New Approach for Discovering Agronomic Important Genes

Besides the natural selection, the crops cultivated today have experienced two episodes of strong artificial selection, domestic and modern breeding. Domestication led to giant genetic structure differentiation between cultivars and their wild species, while modern breeding made further genetic structure differentiation between the modern varieties and the landraces. In a population, diversity of the loci under strong selection is significantly lower than that of other loci. At the same time, diversity in the genomic regions flanking these selected loci also declines in the process of selection. This phenomenon is called hitchhiking effects or selection sweep in genetics. Genomic regions with selection sweep （haplotype block） could be detected after draft genome scanning （genome typing） with molecular markers in a number of released varieties or natural populations. Marker/trait association analysis in these regions would detect the loci （or QTLs） even the favored alleles （genes） in breeding or natural adaptation. Fine scanning of these genomic regions would help to determine the sizes of haplotype blocks and to discover the key genes, thereby providing very valuable information for isolation of the key genes and molecular design of new varieties. Establishment of high density genetic linkage maps in the major crops and availability of high throughput genotyping platform make it possible to discover agronomic important genes through marker/trait association analysis. On the basis of available publications, we give a brief introduction of the hitchhiking effect mapping approach in this paper using plant height, 1 000-grain weight, and phosphorus-deficiency tolerance as examples in wheat.

THE EFFECT OF db-cAMP ON THE GENE EXPRESSION OF CALMODULIN AND CYTOSKELETON IN THE TRANSFORMED CELLS

We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 cells. After treatment with 1 mM db-cAMP for 1 hour and 2 hours, there was an early and repldly reduced in gene expression of Calmodulin and c-fos respectively. After db-cAMP treatment for 4 -5 days, the number of capping cells of ConA binding decreased significantly and the cell surface microvllll decreased as well. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed In G1 phase, we have found that the db- cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, mlcrofilaments and fibronectln were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the Inhibition of proliferation, alteration, of phenotype and reco- very of cytoskeleton is transformed cells after treatment with db-cAMP are related to the Inhibition of gene expression of Calmodulin.

Cloning and expression of human arresten gene and effect of its recombinant protein on endothelial cell proliferation

To clone human arresten gene and investigate biological activity of the recombinant protein.Methods Human arresten gene was obtained from the plasmid pGEMArr and subcloned into the BamHⅠ and Pst Ⅰ restriction sites of prokaryotic expression vector pRSET containing T7 promoter.The recombinant plasmid pRSETAN was subsequently transformed into the strain E.coli BL21(DE3),and the target gene was expressed under induction of IPTG.The expressed protein was extracted,purified by Ni 2+ chelation affinity chromatography and refoled.The effect of the recombinant protein on proliferation of human umbilical vein endothelial cells (HUVECs) was also analyzed with the MTT assay.Results Endonuclease digesting and DNA sequencing confirmed that the arresten gene was correctly inserted into the expression vector.The recombinant protein was hightly expressed in the form of inclusion body in the host bacteria after induction.SDS-PAGE analysis revealed that the recombinant protein with a molecular weight of 26×103 amounted to 27% of the total bacterial proteins.The purity of the expected protein could reach over 96% through affinity chromatography.After renaturation,the recombinant protein could reach over 96% through affinity chromatography.After renaturation,the recombinant protein could significantly suppress proliferation of human umbilical vein endothelia cells(HUVECs) induced by vascular endothelial growth factor(VEGF).Conclusion Human arresten gene was successfully cloned into the expression vector pRSET and expressed at high level in Escherichia coli.Purified and refolded arresten protein could effectively inhibit proliferation of vascular endothelia cells.2 refs.

Research on immune effect of two gene vaccines containing brZPC′and brLDHC_4′when in combined inoculation

ZPC and LDHC_4 play a key role in the process of recognition between sperm and egg or sperm movement,respectively.In this study,partial cDNA sequences of ZPC and LDH-C_4 of Microtus branditi(brZPC and brLDHC_4,respectively) were cloned by RT-PCR,and directly inserted into pCR3.1 vector to construct two gene vaccines(pCR3.1-brZPC' and pCR3.1-brLDHC'_4).pCR3.1-brZPC' and pCR3.1-brLDHC,' could express corresponding proteins in transiently transfected CHO cells.The adult female BALB/c mice were inoculated with the recombinant vaccine alone on in combination.The immunized mouse can produce specific antibody that recognizes the corresponding recombinant protein expressed by BL21 in vitro.Moreover,antibodies produced by combinedly immunized mouse were specific and direct,with no inhibitory effect between two vaccines observed when in combined inoculation.The test of cytokines indicated that the expression of IFNγin pCR3.1-brZPC'- and combinedly inoculated mouse increased obviously and the expression of IL2 in pCR3.l-brLDHC'_4- and combinedly inoculated mouse increased obviously,while the expression of IL4 in pCR3.1-brZPC,pCR3.l-brLDHC'_4 and combinedly inoculated group increased obviously.It was suggested that the combined inoculation with pCR3.1-brZPC' and pCR3.l-brLDHC'_4 could induce both humoral immune response and CTL response.In some sense,the combined inoculation may achieve a better contraceptive effect.The results also showed that,when used alone or in combination,these two recombinant vaccines did not do much harm to the follicular development of immunized mouse.So the combined inoculation with these two recombinant vaccines could be a better way to immunocontraception.This study may provide a theoretical basis for the following tests on antifertility in vivo.

PLASMID VACCINE ENCODING HN GENE FROM NEWCASTLE DISEASE VIRUS HAS MARKED ANTITUMORAL EFFECT IN VITRO

Objective: To explore the antitumor effects of hemaagglutinin-neuraminase gene (HN gene) from Newcastle disease virus. Methods: Plasmid vaccine of pIRHN was constructed and transfected into HeLa cells. The expression of HN was analyzed by Western blot analysis, and the mode of cell death was detected by fluorescence microscope, gel electrophoresis and TUNEL assay and the expression of p53 and bcl-2 was also analyzed in transfected Hela cells. The effect of pIRHN on sialic acid contents in the Hela cell was examined. Results: pIRHN nucleic acid vaccines could be expressed in eukaryotic cell. pIRHN could induce apoptosis after HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells, and there was no prominent evidence for the relatedness of the antitumor effect with the expression of p53 and bcl-2. Conclusion: pIRHN may become a new antitumor biological agent.

Nature of Gene Action and Maternal Effects for Pod Borer, <i>Helicoverpa armigera</i>Resistance and Grain Yield in Chickpea, <i>Cicer arietinum</i>

Information on mechanisms and inheritance of resistance is critical to plan an effective strategy to breed for resistance to insect pests. Therefore, we evaluated a diverse array of chickpea genotypes (eight desi and one kabuli) with varying levels of resistance to the pod borer, Helicoverpa armigera to gain an understanding of the nature of gene action and possible maternal effects. The test genotypes were crossed in all possible combinations for a full diallel. The 72 F1s (36 direct and 36 reciprocal crosses) along with the parents were evaluated for resistance to H. armigera under field conditions, and for antibiosis mechanism of resistance (larval survival and larval weight gain) by using detached leaf assay under laboratory conditions, and grain yield under un-protected conditions in the field. Additive gene action governed the inheritance of resistance to H. armigera, while non-additive type of gene action was predominant for inheritance of antibiosis component of resistance (larval survival and larval weight) and grain yield. Greater magnitude of σ2 A(17.39 and 1.42) than σ2 D (3.93 and 1.21) indicated the preponderance of σ2 Ain inheritance of resistance to pod borer, H. armigera under laboratory and field conditions, respectively. There were no maternal effects for inheritance of resistance to pod borer and grain yield. Lines with significant gca effects for pod borer damage and grain yield were identified for further use in the resistance breeding program. The implications of the inheritance pattern of pod borer resistance and grain yield are discussed in the context of strategies to enhance pod borer resistance and grain yield in chickpea.

Anti-tumor effects induced by gene vaccines co-expressing truncated human prostate specific membrane antigen gene and mouse 4-1BBL

Objective To investigate the influence of m4-1BBL on anti-tumor effects induced by truncated human prostate specific membrane antigen ( tPSMA ) gene in mice. Methods A eukaryotic expression plasmid encoding tPSMA and m4-1BBL ( pDC316-tPSMA-IRES m4-1BBL) ,pDC316-tPSMA and pDC316 were constructed.

Genetic expression and effects of the rolled leaf gene Rl(t) in hybrid rice (Oryza sativa L.)

We constructed a nearisogenic line of rolled leaf gene Rl(t), which ex-pressed incompletely dondnance for the character of rolled leaf(RL), with ge-netic background of Zhenshan 97B. Using RL Zhenshan 97B and the originalZhenshan 97B as the female parents, and Minghui 63 and Yanhui 559 as themale parents, crosses of RL Shanyu 63 (RS63) and Shanyu 63(S63), RLShanyou 559 (RS559) and Shanyou 559 (S559) were made. Inheritance andeffects of Rl(t) in hybrid rice were studied at the flowering and at the 20 d afterflowering, respectively. Results were as follow:

EFFECTS OF PSEUDOFYPE RETROVIRUS CONTAINING HUMAN N-RAS ANTISENSE GENE ON THE GROWTH OF HUMAN LIVER CANCER LTNM4 TRANSPLANTED IN NUDE MICE

An amphotropic pseudotype retrovirus containing human N-ras antisense gene was constructed and packaged with helper cells. It has been previously demonstrated that the virus did inhibit the growth of human hepatocarcinoma cell line PLC PRF/5 in vitro accompanied with the blockage of p21 expression. Based on these results, further study was carried on to examine the effect of these viruses on the growth of human hepatoma transplanted LTNM4 in nude mice. It has been shown that the retrovirus containing human antisense N-ras gene could inhibit the hepatoma in nude mice at a rate of 78% (P<0.05) as compared with saline control. No inhibition was observed in group treated with retrovirus which contained no N-ras sequence. These results in vivo lend further support that human N-ras antisense gene mediated by retrovirus could block the expression of the relevant oncogene and lead to the inhibition of cancer growth. It also provided the basis for further approaches of gene therapy for human cancer.

安罗替尼联合多西他赛治疗驱动基因阴性晚期非小细胞肺癌的临床疗效研究

目的 探究非小细胞肺癌治疗中安罗替尼与多西他赛联合的作用。方法 选取吉林省肿瘤医院于2022年6月—2023年7月收治的120例非小细胞肺癌患者为研究对象,采用随机摸球法分为两组,各60例。对照组使用多西他赛治疗,研究组联用多西他赛和安罗替尼治疗。对比两组临床疗效、上皮特异性黏附分子(Epithelial Cell Adhesion Molecule, EP-CAM)、可溶性细胞间黏附分子-1(Soluble Intercellular Adhesion Molecule-1, SICAM1)、血清细胞黏附分子(Cellular Adhesion Molecule, CAM)水平及不良反应发生率。结果 研究组的疾病缓解率为43.33%,高于对照组的21.67%,差异有统计学意义(χ^(2)=6.419,P=0.011)。治疗后,研究组EP-CAM、SICAM1、CAM水平低于对照组,差异有统计学意义(P均<0.05)。研究组不良反应发生率低于对照组,差异有统计学意义(P<0.05)。结论 安罗替尼联合多西他赛治疗驱动基因阴性非小细胞肺癌能够降低肿瘤标志物,且有助于降低不良反应发生风险。

药物基因检测对难治性精神分裂症患者治疗预后的影响

目的探讨药物基因组学检测结果应用对难治性精神分裂症患者疗效及药物不良反应的影响。方法选取2020年1月-2022年6月江苏省淮安市第三人民医院收治的100例难治性精神分裂症患者。依据基因检测结果指导用药,分别于治疗前、治疗4、8、12、16周使用阳性阴性症状量表(positive and egative symptom scale,PANSS)、临床疗效总评量表(clinical global impression,CGI)评定临床疗效,威斯康星卡片分类测验及个人和社会功能评估量表(personal and social function assessment scales,PSP)分别评定认知及社会功能改善情况,同时使用药物副反应量表(treatment emergent symptom scale,TESS)及做血常规、肝功能、肾功能和心电图等检查,以了解药物不良反应。结果治疗4周PANSS评分为(59.62±6.29)分,治疗8周PANSS评分为(54.83±7.37)分,治疗12周PANSS评分为(49.34±7.93)分,治疗16周PANSS评分(44.68±8.73)分,均低于治疗前的(62.93±5.55)分(P<0.001);治疗4、8、12和16周的CGI、PSP、威斯康星卡片分类测验等评分均优于治疗前(P<0.001)。治疗16周TESS评定与治疗4周比较,差异有统计学意义(P<0.01),但血常规、心电图、脑电图、肝功能和肾功能检查异常与否与治疗前比较,差异无统计学意义(P>0.05)。结论应用基因检测可显著提高难治性精神分裂症患者的临床疗效,且并不增加不良反应,因此基因检测可促进该病的临床合理用药、精准用药和个体化治疗。

茶树炭疽病菌拮抗链霉菌的筛选及其抑菌特性研究

茶炭疽病菌刺盘孢菌(Colletotrichum camelliae)是引起茶树炭疽病的重要致病菌,为获得对C.camelliae具有拮抗作用的链霉菌,采用稀释涂布法和平板对峙法从茶园生境中分离筛选出对其具有明显拮抗作用的链霉菌菌株,并结合形态学观察、生理生化特征和16 SrRNA基因序列分析对其进行种属鉴定;开展基于链霉菌菌株的抗菌谱测定、茶炭疽病菌菌丝生长抑制试验和孢子萌发抑制试验;通过菌丝生长速率法测定其无菌发酵滤液对茶炭疽病菌的抑菌活性及其抑菌活性稳定性,并测定其产胞外降解酶能力、抗菌物质合成基因、挥发性与非挥发性代谢物抑菌活性。结果表明,筛选获得一株对茶炭疽病菌C.camelliae具有良好抑菌效果的菌株XS-4,对茶炭疽病菌的平板抑制效果为76.42%;结合形态学观察、生理生化特征,以及16 SrRNA基因序列分析,将菌株XS-4鉴定为多产色链霉菌(Streptomyces polychromogenes);菌株XS-4对其他8种植物病原菌均具有较好的抑菌效果,抗菌性能具有广谱性。扫描电镜结果表明,菌株XS-4能抑制茶炭疽病菌菌丝生长。孢子萌发抑制试验发现,拮抗菌XS-4发酵液能抑制茶炭疽病菌的孢子萌发,抑制率为62.48%;菌株XS-4的最佳发酵培养基为KMB培养基,在KMB培养基中培养7 d所产生的抑菌活性物质对茶炭疽病菌的抑菌效果最好;其无菌发酵滤液对温度、酸碱度、紫外照射、蛋白酶均具有较好的稳定性;菌株XS-4的非挥发性代谢物对茶炭疽病菌的抑菌活性较好,抑菌率达81.92%;菌株XS-4具有产生淀粉水解酶、蛋白酶、β-1,3-葡聚糖酶、纤维素酶的能力;菌株XS-4具有产生抗菌物质的pks-Ⅰ及pks-Ⅱ基因。综上所述,菌株XS-4在茶树炭疽病生物防治方面具有巨大的应用潜力。