Genotypes Analyses of Ten Genes Related to Eating Quality of Twenty-three Fragrant Rice Cultivars by Molecular Marker

Fragrance is a very important quality traits of rice, fragrant rice research has been an important research issue in rice research fields in nowadays. In our study, using the detection method of molecular markers, genotypes of the 10 genes (Wx, SSII 3, SBE3, AGPiso, SSIII 2, AGPlar, PUL, SSI, ISA, SSIV 2) associated rice eating quality were analyzed in the 23 fragrant rice cultivars. The result indicated that the “2845”rice cultivar possesses the best combination of genotypes related to the highest eating quality, and the next best rice cultivars were “Songxiangzaojing”, ”Suhuxiangjing”, “B1”, “Wuyun2645”, “Tongyunjing”, “Yinxiang28”, “Xiangjing49②”, “99983”, “Wxiang99075”, “07-08”, “Yunjingyou15”, “29185”, “Nanhai318”. Other 9 rice cultivars (“Dahuaxiangjing”, “Wuxiang14”, “Xiangjing”, “Della”, “Dalixiang”, “Taiguoxiangdao”, “Cxiang517”, “Xiangdao No1”, “Zhongxiang No1”) obtain some of bad genotypes which may have adverse effect on rice eating quality. This study was helpful to establish an overall understanding of the genotypes of these ten eating quality related genes in the 23 tested fragrant rice cultivars, and provides the important genotype information in selecting and cultivating new fragrant rice with good eating quality in future via molecular markers assisted selection method.     

雪花莲外源凝集素基因转化番茄

The plasmid pRSSGNA1 carried a snowdrop lectin gene (Galanthus nivalis agglutinin, GNA) under the drive of RSs-1 promoter, were successfully transferred into three tomato (Lycopersicon esculentum Mill.) cultivars, “C8”,“A39” and “A53” via Agrobacterium-mediated transformation. Regenerated plantlets were obtained from cotyledons after preculture, shoot inducing culture and root inducing culture. Transgenic tomato plants were confirmed by the kanamycin-resistant experiment, PCR analysis and Southern blot. The preliminary results from bioassay demonstrated significant resistance of the transgenic plants to aphid (Myzus persicae Sulzer) larvae. The inheritance of selective marker gene (NPTⅡ) in 3 transgenic tomato plants is in the model of the simple Mendel's fashion in progenies of the selfing generation.     

Induction of a β-phaseolin promoter by exogenous abscisic acid in tobacco: developmental regulation and modulation by external sucrose and Ca2+ ions

Using degenerate oligonucleotides that correspond to conserved amino acid residues of known 1-aminocyclopropane-1-carboxylic acid (ACC) synthases, we cloned a genomic fragment that encodes ACC synthase in Stellaria longipes. Southern analysis suggests that ACC synthase is encoded by a small gene family comprising about 4 members. We isolated four unique ACC synthase cDNA clones under different growth conditions from alpine and prairie ecotypes of S. longipes. Northern analyses suggest that ACC synthase genes are differentially and synergistically regulated by photoperiod and temperature. Such differential regulation of ACC synthase genes positively correlate with the levels of ACC and ethylene. Since ethylene has previously been shown to partly control the stem elongation plasticity in S. longipes, we propose that differential regulation of ACC synthase genes may represent one of the underlying molecular mechanisms of phenotypic plasticity in S. longipes.     

Effect of Heat Stress on Photosynthetic Characteristics of Different Green Organs of Winter Wheat During Grain-filling Stage

Four winter wheat ( Triticum aestivum L.) varieties (“JD 8”, “Jing 411”,“Centurk” and “Tam 202”) were used to study the effect of heat stress on photosynthetic characteristics of flag leaf blade, flag leaf sheath, peduncle, glume, lemma and awn during grain-filling stage. The results showed that heat acclimation during grain-filling stage increased thermotolerance of wheat with significant differences among different green organs. During heat stress, the decreases of the efficiency of primary light energy conversion ( Fv/Fm ) of PS and pigment (chlorophyll and carotenoid) content were much slower in peduncle, flag leaf sheath and glume than in flag leaf blade, lemma and awn; and the percentage of decrease in net photosynthetic rate ( Pn ) of ear was lower than that of the flag leaf blade. The measured photosynthetic parameters ( Fv/Fm , Pn and pigment content) of “JD 8”, a relatively heat tolerant variety, declined more slowly than those of the other three varieties during the whole heat stress period.      

水稻的光合作用、生长、碳同位素分辨作用及抗渗透胁迫性对CO2浓度增高的响应

四个水稻(Oryza sativa L.)品种"IR72"、"特三矮2号"、"桂朝2号"和"Ⅱ优4480"在田间栽于含35 μmol/mol 和60 μmol/mol CO2的塑料大棚中,自然光照.高浓度CO2下供试水稻品种的光合速率变化表现为提高型("IR72"、"特三矮2号")、稳定型("桂朝2号"的Pn几无变化)和下调型("Ⅱ优4480").生长速率、穗重、由Δ13C计算而得的长期水分利用效率和清除DPPH@自由基的能力皆增加.除"Ⅱ优4480"外,其他3个品种明显增高总生物量.供试品种的穗重/总生物量比不同程度地受到高浓度CO2的改变.叶片段经PEG渗透胁迫后,不同的生长于高浓度CO2者的电解质渗漏率较小.结果表明高浓度CO2可改变水稻的光合作用和水分关系特性,品种间不同的响应显示了选育适于未来高浓度CO2下具有高产和抗逆性品种的可能性.     

水稻基因APRT的克隆及其与温敏核雄性不育的关系

在拟南芥中腺嘌呤磷酸核糖转移酶基因(APRT)突变导致植株雄性不育.本文首次报道从水稻(Oryza sativa subsp.indica)中克隆了基因APRT(GenBank登录号AY238894),并将其定位于水稻第4染色体的一个BAC克隆(AL606604)的58 000 bp至63 000 bp区域.该基因长4 220 bp(起始密码子至终止密码子),含7个外显子、6个内含子,编码的APRT蛋白长212个氨基酸残基,与其他物种来源的APRT序列存在很高的同源性.与大麦、小麦、拟南芥1型及其2型的该蛋白同源性分别为54.9%、54.9%、49.6%和59.5%.经保守结构域搜索发现该蛋白中存在APRT催化结构域.从DNA、mRNA两个水平分析了该基因与水稻温敏核雄性不育(TGMS)的关系,结果表明:受温度诱导,水稻"安农S-1"APRT基因的表达变化可能与温敏核雄性不育表现型具相关性.     

Identification of the Molecular Markers Linked to the Salt-resistance Locus in the Wheat Using RAPD-BSA Technique

Calli from mature embryo of “Jimai-24” wheat ( Triticum aestivum L.) were induced on medium containing Zhengdingmycin then continuously cultured on medium containing 0.5% NaCl till to regenerate plants named 8901-17 salt-tolerant mutant. “Jimai-24” was compared with 8901-17 by using the technique of RAPD. Thirty-five out of 280 random primers could detect DNA polymorphism. The similarity index was 0.978, indicating that they were NILs (near-isogenic lines). Two F2 populations (“Jimai-24”×8901-17 and 8901-17דZhongmai-9”) had been constructed using the method of half-division. The two relative DNA pools (salt tolerant DNA pool and susceptible DNA pool) which come from the two F2 populations, respectively, had been made according to the method of BSA (bulked segregant analysis). RAPD analysis between the two DNA relative pools was carried on with above 35 random primers which could detect DNA polymorphism definitely. The identical polymorphism between the two sets of DNA pools come from the two F2 populations could be determined only by OperonQ4 primer. This result implied that the polymorphic fragment amplified by OperonQ4 primer was the molecular marker of RAPD closely linked to the salt tolerant mutation locus.     

与授粉有关的因子调节的乙烯生物合成基因在兰花花器官中的表达

分析了与授粉有关的因子调节的ＡＣＣ合酶和ＡＣＣ氧化酶基因在朵丽蝶兰（ＤｏｒｉｔａｅｎｏｐｓｉｓｈｙｂｒｉｄａＨｏｒｔ．）花中的表达。生长素和乙烯均可诱导ＡＣＣ合酶和ＡＣＣ氧化酶的ｍＲＮＡ在花器官中积累。然而,去雄却不能诱导这两个基因在花器官中表达。生长素和乙烯所诱导的ＡＣＣ合酶和ＡＣＣ氧化酶的ｍＲＮＡ在花器官中的积累模式相似。原位杂交结果表明,生长素和乙烯处理后ＡＣＣ氧化酶的ｍＲＮＡ在柱头的表皮和薄壁细胞中积累。根据ＡＣＣ合酶和ＡＣＣ氧化酶基因表达的结果,对生长素、乙烯和去雄在兰花授粉后乙烯生物合成过程中的作用进行了分析。     

Comparative Spectroscopic Studies on the Photoinhibition Process in Photosystem ￠?Complex from Two Wheat Cultivars

The photodamage processes of PSⅠ particles isolated from two wheat cultivars “Jing 411” and “Xiaoyan 54” were studied by comparing the difference in spectroscopic properties. It was found that high light intensity caused the damage of pigments in PSⅠ, especially Chl a molecules with maximum absorption at 683 nm is very sensitive to high light. The change in fluorescence spectra revealed that photodamage also led to the damage of the process of energy transfer in PSⅠ. In the PSⅠ particles “Xiaoyan 54”, the absorption of Chl a molecules at 683 nm slightly decreased at the beginning of illumination and meanwhile the fluorescence become stronger, but the absorption become stable rather long, and declining after 40 min. On the other hand, PSⅠ particles of “Jing 411” showed no such changes during the process of photodamage. Presumably in PSⅠ of “Xiaoyan 54”, excessive energy was distributed to long wave chlorophyll molecules and the number of antenna pigment molecules was less, so that less energy was transferred to the reaction center P700 and thus it was protected. This is the possible reason why “Xiaoyan 54” was more resistant to photooxidation.     

河套蜜瓜ACC合成酶cDNA片段的克隆和序列分析

1-氨基环丙烷-1-羧酸（ACC）合成酶是高等植物中乙烯生物合成的关键酶。以成熟河套蜜瓜（CucumismeloL．cvHetau）果实的RNA为模板,经反转录和PCR扩增得到预期大小的DNA片段,插入到pUC19的SmaⅠ位点后转化E.coliJM109,筛选出重组子pHMAS1。序列分析表明获得了长627bp的ACC合成酶cDNA片段。与已报道的ACC合成酶基因相应序列比较有很高的同源性．     

C4 Pathway Enzymes in Soybean Leaves

High-yielding soybean ( Glycine max (L.) Merr.) variety, “Heinong 40”, and one control variety, “Heinong 37”, were used as experimental materials. The activities of C4 pathway enzymes, i.e. PEPCase (phosphoenolpyruvate carboxylase), NADP-ME (NADP-malic enzyme), NADP-MDH (NADP-malate dehydrogenase), PPDK (pyruvate phosphate dikinase) and RuBPCase (ribulose-1.5-bisphosphate carbozylase) were assayed at seedling, blooming, pod-bearing and pod-filling stage. The results indicated that C4 pathway enzymes were expressed differently at different developing stages in both varieties of soybean, but the ratio of PEPCase and RuBPCase showed that the expression of C4 pathway enzymes of “Heinong 40” was higher than those of “Heinong 37” at each stage. The results showed that C4 pathway enzymes were closely related to the crop yielding potential. Therefore, it is possible to select potentially high-yielding soybean variety by the expression of C4 pathway enzymes.     

钼、硼对大豆叶片膜脂过氧化及体内保护系统的影响

The study deals with the changes of membrane peroxidation and endogenous protective system with different supplementation of molybdenum (Mo) and/or boron (B) concentration in soybean (Glycine max L.) leaves at three developmental stages (5-trifoliate stage, initiation of flowering, and peak podsetting stage) in three pot-grown soybean varieties (“Zhechun No.3”, “Zhechun No.2”, “3811”). The control plants under low Mo and low B exhibited an increasing of membrane permeability (MP), malondialdehyde (MDA) and proline (Pro) contents, polyphenol oxidase (PPO) and ascorbate oxidase (AO) activities and a decrease of ascorbate (AsA) contents, superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (AP) and catalase (CAT) activities. Application of Mo or B alone raised the ability of anti-oxidant of soybean leaves. The results indicated that the anti-oxidant enzymes (including SOD, POD, CAT and AP) related closely to anti-oxidant compounds (including AsA and Pro). There was some difference between the effects of Mo and B on the anti-oxidant, and a synergetic effect was observed between Mo and B. Some genetic variation in the responses to Mo and B was found among the three soybean varieties which was related to the activities of the total anti-oxidant systems.     

Identification of a 1-aminocyclopropane-1-carboxylic acid synthase gene linked to the female (F) locus that enhances female sex expression in cucumber.

Sex determination in cucumber (Cucumis sativus L.) is controlled largely by three genes: F, m, and a. The F and m loci interact to produce monoecious (M_f_) or gynoecious (M_f_) sex phenotypes. Ethylene and factors that induce ethylene biosynthesis, such as 1-aminocyclopropane-1-carboxylate (ACC) and auxin, also enhance female sex expression. A genomic sequence (CS-ACS1) encoding ACC synthase was amplified from genomic DNA by a polymerase chain reaction using degenerate oligonucleotide primers. Expression of CS-ACS1 is induced by auxin, but not by ACC, in wounded and intact shoot apices. Southern blo hybridization analysis of near-isogenic gynoecious (MMFF) and monoecious (MMff) lines derived from divers genetic backgrounds revealed the existence of an additional ACC synthase (CS-ACS1G) genomic sequence in the gynoecious lines. Sex phenotype analysis of a segregating F2 population detected a 100% correlation between the CS-ACS1G marker and the presence of the F locus. The CS-ACS1G gene is located in linkage group B coincident with the F locus, and in the population tested there was no recombination between the CS-ACS1G gene and the F locus. Collectively, these data suggest that CS-ACS1G is closely linked to the F locus and may play a pivotal role in the determination of sex in cucumber flowers.     

Construction of EMS-Induced Peanut Mutant Libraries and Identification of Pod-Related Traits Mutant Lines

Peanut (Arachis hypogaea L.) is an oil and economic crop of vital importance, and peanut pod is the key organ influencing the yield and processing quality. Hence, the Pod-related traits (PRTs) are considered as important agronomic traits in peanut breeding. To broaden the variability of PRTs in current peanut germplasms, three elite peanut cultivars were used to construct Ethyl methane sulfonate (EMS)-induced mutant libraries in this study. The optimal EMS treatment conditions for the three peanut varieties were determined. It was found that the median lethal dose (LD50) of EMS treatment varied greatly among different genotypes. Finally, the EMS-induced peanut mutant libraries were constructed and a total of 124 mutant lines for PRTs were identified and evaluated. Furthermore, “M-8070”, one of the mutant lines for pod constriction, was re-sequenced via high-throughput sequencing technology. The genome-wide variations between “M-8070” and its wild parent “Fuhua 8” (FH 8) were detected. 2994 EMS-induced single nucleotide polymorphisms (SNPs) and 1188 insertion-deletions (InDels) between “M-8070” and its wild parent were identified. The predominant SNP mutation type was C/G to T/A transitions, while the predominant InDel mutation type was “1-bp”. We analyzed the distribution of identified mutations and annotated their functions. Most of the mutations (91.68% of the SNPs and 77.69% of the InDels) were located in the intergenic region. 72 SNPs were identified in the exonic region, leading to 27 synonymous, 43 non-synonymous and 2 stop-gain variation for gene structure. 13 Indels were identified in the exonic region, leading to 4 frame-shift, 8 non-frame-shift and 1 stop-gain variations of genes. These mutations may lead to the phenotypic variation of “M-8070”. Our study provided valuable resources for peanut improvement and functional genomic research.     

一个新的水稻小穗梗弯曲突变体的形态特征及基因定位

稻穗小穗梗的发育与产量有着密切关系。文章利用⁶⁰Co g 射线辐照籼稻品种“浙农7号”, 获得一个性状能稳定遗传的小穗梗弯曲突变体bpb1 (bent pedicel branch 1), 表现为小穗梗弯曲, 并伴有小穗梗长度增长、穗长缩短和植株矮化等特点。扫描电镜观察显示, bpb1突变体小穗梗的表皮毛及气孔变小, 外表皮细胞和厚壁细胞排列不规则, 接近弯曲部位的细胞变小、排列更为紧密。bpb1突变体小穗梗横切面观察表明, 小维管束排列结构发生明显变化。遗传分析表明该突变表型受隐性单基因控制。利用bpb1突变体与粳稻品种“浙农大104”杂交构建的F₂群体进行基因定位, 将bpb1基因定位于水稻第7号染色体长臂SSR标记RM21537和RM21552之间, 该区间的物理距离为343 kb, 该区域内未发现与水稻小穗梗发育相关的已知基因。文章为bpb1基因的克隆和功能研究奠定了重要基础。