Improved bioethanol production using CRISPR/Cas9 to disrupt the ADH2 gene in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Bioethanol, as a form of renewable and clean energy, has become increasingly important to the energy supply. One major obstacle in ethanol production is developing a high-capacity system. Existing approaches for regulating the ethanol production pathway are relatively insufficient, with nonspecific genetic manipulation. Here, we used CRISPR/Cas9 technology to disrupt the alcohol dehydrogenase (ADH) 2 gene via complete deletion of the gene and introduction of a frameshift mutation in the ADH2 locus. Sequencing demonstrated the accurate knockout of the target gene with 91.4% and near 100% targeting efficiency. We also utilized genome resequencing to validate the mutations in the ADH2 mutants targeted by various single-guide RNAs. This extensive analysis indicated the mutations in the CRISPR/Cas9-engineered strains were homozygous. We applied the engineered Saccharomyces cerevisiae strains for bioethanol production. Results showed that the ethanol yield improved by up to 74.7% compared with the yield obtained using the native strain. This work illustrates the applicability of this highly efficient and specific genome engineering approach to promote the improvement of bioethanol production in S. cerevisiae via metabolic engineering. Importantly, this study is the first report of the disruption of a target gene, ADH2, in S. cerevisiae using CRISPR/Cas9 technology to improve bioethanol yield.     

黄羊（Procapra gutturosa）在中国分布的变迁及其资源持续利用

历史上黄羊曾广布于中国的北部,目前黄羊只分布在我国内蒙古北部的中蒙边境一带。环境的变迁、人类活动的干扰及盲目捕猎是黄羊分布区缩小的重要原因。建议尽快建立黄羊自然保护区,使黄羊在我国有一安歇、繁衍的良好栖息地,最终使黄羊资源得到科学合理的利用。     

兰花蕉种子的解剖学和组织化学研究

兰花蕉种子球形或近球形,具表皮毛,种脊不明显。种子包括假种皮、种皮、外胚乳、内胚乳和胚五部分。假种皮具３～４条粗毛状裂片,包围种子或不定向伸展;裂片最外方为１层表皮细胞和１～３层厚壁细胞,内方为薄壁细胞;表皮细胞和厚壁细胞的壁增厚并木质化;成熟时裂片下部１／２段中空。种皮由外珠被发育而来,但内珠被在种子发育后期才萎缩。种皮分化为外种皮,中种皮与内种皮;外种皮由１层表皮细胞构成,其细胞壁增厚并木质化;中种皮外方为２～３层厚壁细胞,内方由１２～１５层薄壁细胞构成;内种皮由１层径向延长的石细胞构成,其细胞壁网状增厚,胞腔不明显。外胚乳极不显眼,大部分只由１层切向延长的长方形细胞构成,局部为２～１７层细胞;外胚乳细胞主要含许多脂类物质及少量蛋白质颗粒,不含淀粉。内胚乳占据种子很大的体积,由通常径向延长的长方形、长条形或方形薄壁细胞构成;细胞内充满淀粉粒和通常一颗亦有２至多颗菱形或方形蛋白质晶体,脂类物质极少。胚圆柱形,胚根和胚芽不明显。种子珠孔区不分化出珠孔领和孔盖,但具柄,柄的远轴端边缘大部分着生假种皮,着生假种皮一侧柄略膨大。合点区内种皮出现极宽的缺口,缺口间为整体呈弧状长条形的合点区厚壁细胞群。较粗的种脊维管?     

Cross-Cultural Validation of the High Blood Pressure Health Literacy Scale in a Chinese Community

BackgroundConsidering the importance of health literacy (HL) for the maximum yield from the hypertension control programs, development of a reliable and valid instrument of hypertension-related HL is critical. This study aimed to translate and validate the High Blood Pressure-Health Literacy Scale (HBP-HLS) into Chinese (C-HBP-HLS) and evaluate its psychometric properties in Chinese context.MethodBetween June 2013 and January 2014, a cross-sectional study was conducted among recruited hypertensive patients belonging to the Han and Kazakh-Chinese communities in Urumqi, Xinjiang, China.ResultsA pilot sample (n = 242) was selected for the exploratory factor analysis of the translated and modified instrument. Another sample (n = 308) was recruited for the confirmatory factor analysis. C-HBP-HLS consisted of five dimensions (Print Health Literacy, Medication Label, Understanding Ability, Newest Vital Sign Test, and Avoiding Food Allergy) containing 15 items, accounting for 77.7% of the total variance. The 5-factor model demonstrated a good overall fit. The scale-level content validity index was 0.85. Cronbach’s alpha of the overall scale was 0.78 and test-retest reliability was 0.96. Education level had a strong positive correlation with the scores for items Q1, Q2, and Q3(r = 0.481, 0.492, 0.475, respectively). Health Literacy scores among Kazakh patients were significantly lower than Han (7.13±7.90 vs. 30.10±13.42, Z = -14.573, P<0.001).ConclusionC-HBP-HLS demonstrated suitable factor structure and robust psychometric properties for measuring health literacy level among hypertensive patients in China.     

Analysis of Xyloglucan Endotransglycosylase/Hydrolase (XTH) Genes and Diverse Roles of Isoenzymes during Persimmon Fruit Development and Postharvest Softening

Xyloglucan endotransglycosylase/hydrolase (XTH) enzymes have played a role in the remodeling of cell wall hemicelluloses. To investigate the function of XTHs in persimmon (Diospyros kaki L.) fruit development and postharvest softening, five cDNAs (DkXTH1 to DkXTH5), whose putative proteins contained the conserved DEIDFEFLG motif of XTH, were cloned. Real time quantitative PCR analysis revealed that DkXTH1, DkXTH4, and DkXTH5 peaked in immature expanding fruit, and their higher expression was observed along with higher fruit firmness in cold-treated fruit or firmer cultivar fruit during storage. The opposite gene expression patterns were observed in DkXTH2 and DkXTH3, which reached maxima concomitance with pronounced fruit softening. Meanwhile, the xyloglucan endotransglycosylase (XET) enzymes play important roles in both the rapid growth and ripening of persimmon fruit. Furthermore, the recombined DkXTH1 and DkXTH2 proteins showed significant XET activity without any detected XEH activity. However, the XET activity of recombined DkXTH2 protein had a higher affinity for small acceptor molecules than that of recombined DkXTH1 protein. The former might prefer to participate in cell wall restructuring, and the latter is more inclined to participate in cell wall assembly. Besides, DKXTH proteins could function by targeting to the cell wall under regulation of a signal peptide. The data suggested that individual DKXTHs could exhibit different patterns of expression, and the encoded products possessed specific enzymatic properties conferring on their respective functions in growth and postharvest softening of persimmon fruit.     

Prevalence and risk factors of overweight and obesity in China

Objective: To examine the prevalence and risk factors of overweight and obesity in China. Research Methods and Procedures: A cross‐sectional survey was conducted in a nationally representative sample of 15,540 Chinese adults in 2000–2001. Body weight, height, and waist circumference were measured by trained observers. Overweight and obesity were defined according to the World Health Organization classification. Central obesity was defined according to guidelines of the International Diabetes Federation. Results: Mean BMI and waist circumference were 23.1 kg/m² and 79.6 cm, respectively, for men and 23.5 kg/m² and 77.2 cm, respectively, for women. The prevalences of overweight and obesity were 24.1% and 2.8% in men and 26.1% and 5.0% in women, respectively. The prevalence of central obesity was 16.1% in men and 37.6% in women. The prevalences of overweight, obesity, and central obesity were higher among residents in northern China compared with their counterparts in southern China and among those in urban areas compared with those in rural areas. Lifestyle factors were the most important risk factors to explain the differences in overweight and central obesity between northern and southern residents. Among women, lifestyle and diet were the most important risk factors to explain the differences between urban and rural residents, whereas socioeconomic status, lifestyle, and diet were all important among men. Discussion: Our study indicates that overweight and obesity have become important public health problems in China. Environmental risk factors may be the main reason for regional differences in the prevalence of overweight and obesity in China.     

云南曲靖志留纪罗德洛世真盔甲鱼类一新属(英文)

记述了云南曲靖潇湘区关底组(志留系罗德洛统)真盔甲鱼类一新属、新种——长孔盾鱼(Dunyu longiforus gen.et sp.nov.),丰富了以梦幻鬼鱼为代表的潇湘脊椎动物群的组成。该新属具真盔甲鱼科的鉴别特征:后眶上管和中背管二者连续过渡,后端两侧辏合呈"U"字形,侧背管仅发出3条侧横管。新属内角缺如,这一特征过去在真盔甲鱼科中仅见于真盔甲鱼属。新属区别于真盔甲鱼属的主要特征有:角向后方延伸,呈棘状或叶状;头甲最宽处位于角末端之前,宽长比小于1.1;中背孔末端超越眶孔后缘连线。基于对秀山真盔甲鱼头甲标本的重新观察,将秀山真盔甲鱼归入盾鱼属。新种区别于秀山盾鱼(Dunyu xiushanensis)的特征有:个体较大,头甲长大于宽;角呈棘状;眶孔位置相对靠前,头甲眶前区与眶后区的长度之比小于0.75;多边形崮状瘤点,较大,长可超过2mm。     

神香草的组织培养及植株再生

1植物名称神香草(HyssopusofficinalisLinn.)。2材料类别嫩茎。3培养条件培养基:(1)MS+6-BA0.2mg·L-1(单位下同)+NAA0.2+La(NO3)31.0;(2)MS+6-BA0.4+NAA0.4+La(NO3)31.2;(3)MS+6-BA0.6+NAA0.6+La(NO3)31.4;(4)MS+6-BA0.8+NAA0.8+La(NO3)31.6;(5)MS+6-BA0.8+NAA0.8+La(NO3)31.8;(6)MS+6-BA0.2+IAA0.2;(7)MS+6-BA0.4+IAA0.4;(8)MS+6-BA0.6+IAA0.6;(9)MS+6-BA0.8+IAA0.8;(10)MS+6-BA1+NAA1;(11)MS+6-BA1.2+NAA1.2;(12)MS+6-BA0.1+IAA0.6;(13)1/3MS+IAA0.5;(14)1/3MS+IAA1.2。上述MS培养基中加3…