许多蛾类的两性化学通信通常具有显著的地理差异性，因此进一步明确特定地理种群的性信息素系统有益于提高害虫防治效果。本研究系统分析了中国新疆地区亚洲玉米螟种群的性信息素产生与行为反应。已知该雌蛾性腺中可合成(Z)-12-tetradecenyl acetate (Z12-14:Ac)，(E)-12-tetradecenyl acetate (E12-14:Ac)和n-tetradecyl acetate (14:Ac)三种主要成分，且三组分的比例呈现显著的地理性差异。前两种组分一般认为是该物种的性信息素，而14:Ac在不同地理种群中的行为调控功能不同。对新疆种群的性信息素鉴定定量分析表明雌蛾腺体中Z12-14:Ac与E12-14:Ac的平均比例以及14:Ac在三组分中的平均占比分别为60.46% (SD=5.26) 和25.00% (SD=7.37)，且两种比例数据呈正态或近正态分布。玉米田中的诱捕试验表明诱芯中添加近似腺体比例的双组分(Z12-14:Ac和E12-14:Ac)与三组分诱捕量均最大，而偏离腺体比例诱捕量均降低。最佳比例三组分的诱捕量显著高于最佳比例的双组分，因此14:Ac为本地种群的性信息素成分。此外，诱芯中最佳比例的三组分剂量在200 到 350 μg之间活性较高。本研究为亚洲玉米螟性信息素通信的地理差异提供了更多信息，并为提高新疆种群的性信息素监测与防治效果提供依据。

外源基因在转基因动物和细胞中稳定、高效的表达，对于基因功能的研究和生物反应器的建立至关重要。动物基因组中的友好基因座能使外源基因高效稳定的表达且无副作用，但目前猪基因组中可供外源基因安全、高效定点整合的基因座相对较少，限制了多转基因猪的研究和发展。本研究提出了一种将猪I型胶原α1链（collagen type I alpha 1 chain，COL1A1）基因座作为友好基因座表达外源基因的新策略。利用CRISPOR软件设计了一对靶向COL1A1基因终止密码子的sgRNA，并连接到CRISPR/Cas9表达载体pX330中；同时合成了一个不含启动子、左右同源臂各长为900 bp的2A-GFP供体载体，然后共转染猪肾上皮细胞（porcine kidney epithelial，PK15)，胎儿成纤维细胞（porcine embryonic fibroblast，PEF）以及回肠上皮细胞（porcine intestinal epithelial，IPI-2I）3种细胞。电转染48 h后观察3种细胞的荧光情况，72 h后通过流式细胞术分选绿色荧光蛋白阳性的细胞，并通过荧光定量、蛋白质印记、转录组测序和CCK8实验在不同水平上评估COL1A1基因作为猪基因组外源基因定点整合位点的安全性。结果表明，共转染sgRNA和供体载体之后，可以分别在猪3种不同细胞中观察到绿色荧光，说明同源重组介导的定点整合系统可以在猪基因组中很好的发挥作用。qPCR结果显示，GFP敲入对3种细胞COL1A1基因mRNA的表达并无显著影响（PK15、PEF和IPI-2I细胞中的P值分别为0.29、0.66和0.20）。同样，蛋白质印记结果显示，GFP阳性细胞与野生型细胞COL1A1蛋白的表达并无显著差异（PK15、PEF和IPI-2I细胞中的P值分别为0.64、0.48和0.80)。转录组测序结果显示，GFP阳性PEF细胞与野生型PEF细胞的转录组显著正相关（P<2.2e-16），表明GFP敲入没有改变内源性基因的整体表达。CCK8实验表明，GFP敲入对PK15细胞增殖并无显著影响（24 h，48 h，72 h，96 h，120 h的P值分别为0.31，0.96，0.24，0.17和0.38）。上述结果表明，COL1A1基因可作为猪基因组中外源基因定点整合的友好基因座，可广泛应用于家畜育种和生物医学模型的建立。本研究首次鉴定出了COL1A1基因可以作为猪基因组中的友好基因座。

小麦抗旱相关基因TaPYL4的功能分析及标记开发

坡耕地氮素流失是农业面源污染的重要原因之一，严重影响着水环境。本研究以顺坡垄作、横坡垄作坡面为研究对象，通过两年(2017-2018年)野外定外观测，开展玉米生长期黄壤坡耕地地表径流、壤中流及其氮素流失特征。研究结果表明：随着降雨量的增加，地表径流量和壤中流量也随之增加。顺坡垄作的地表径流量和氮素流失量均显著高于横坡垄作；横坡垄作0 -20 cm和20 -40 cm壤中流量和氮素流失量均显著高于顺坡垄作。地表径流的总氮流失量占径流总氮流失的54.95 -81.25%，因此，我们推断地表径流是研究区氮素流失的主要途径。不同耕作措施下可溶性总氮是氮素流失的主要形式，其流失量占氮素流失量的55.82-94.41%，而可溶性有机氮占可溶性总氮流失量的52.81-87.06%。因此，我们推断可溶性氮为研究区氮素流失的主要形式。应在玉米苗期，进一步开展氮素流失防控方面研究，以减少通过径流途径铵态氮流失造成的环境污染。

Starch is the most important component in endosperm of sorghum grain.  Usually, two types of starch are present: amylose (AM) and amylopectin (AP).  The levels of AM and AP contents play a significant role in the appearance, structure, and quality of sorghum grains and in marketing applications.  In the present study, a panel of 634 sorghum (Sorghum bicolor (L.) Moench) accessions were evaluated for starch, AM, and AP contents of grain, which included a mini core collection of 242 accessions from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) in India, and 252 landraces and 140 cultivars from China.  The average starch content was 67.64% and the average AM and AP contents were 20.19 and 79.81%, respectively.  We developed a total of 260 000 high-confidence single nucleotide polymorphism (SNP) markers in the panel of 634 accessions of S. bicolor using specific locus amplified fragment sequencing (SLAF-seq).  We performed genome-wide association studies (GWAS) of starch, AM, and AM/AP of grain and SNP markers based on a mixed linear model (MLM).  In total, 70 significant association signals were detected for starch, AM, and AM/AP ratio of grain with P<4.452×10^–7, of which 10 SNPs were identified with significant starch, 51 SNPs were associated with AM, and nine SNPs were associated with the AM/AP ratio.  The Gene Ontology (GO) analysis identified 12 candidate genes at five QTLs associated with starch metabolism within the 200-kb intervals, located on chromosomes 1, 5, 6, and 9.  Of these genes, Sobic.006G036500.1 encodes peptidyl-prolyl cis-trans-isomerase CYP38 responsible for hexose monophosphate shunt (HMS) and Sobic.009G071800 encodes 6-phospho-fructokinase (PFK), which is involved in the embden-meyerhof pathway (EMP).  Kompetitive allele specific PCR (KASP) markers were developed to validate the GWAS results.  The C allele is correlated with a high starch content, while the T allele is linked with a low level of starch content, and provides reliable haplotypes for MAS in sorghum quality improvement.

Cotton plants are recalcitrant with regards to transformation and induced regeneration.  In the present study, 5-enolpyruvylshikimate-3-phosphate (EPSPS), a glyphosate resistant gene from the bacterium Agrobacterium sp. strain CP4, was introduced into an elite Bt transgenic cotton cultivar with a modified technique involving in planta Agrobacterium-mediated transformation of shoot apex.  Primary transformants were initially screened using a 0.26% glyphosate spray and subsequently by PCR analysis.  Five out of 4 000 transformants from T₁ seeds were obtained resulting in an in planta transformation rate of 0.125%.  Four homozygous lines were produced by continuous self-fertilization and both PCR-based selection and glyphosate resistance.  Transgene insertion was analyzed by Southern blot analysis.  Gene transcription and protein expression levels in the transgenic cotton lines were further investigated by RT-PCR, Western blot, and ELISA methods.  Transgenic T₃ plants were resistant to as much as 0.4% of glyphosate treatments in field trials.  Our results indicate that the cotton shoot apex transformation technique which is both tissue-culture and genotype-independent would enable the exploitation of transgene technology in different cotton cultivars.  Since this method does not require sterile conditions, the use of specialized growth media or the application of plant hormones, it can be conducted under the greenhouse condition.

Radiation sensitivity proteins-23 (RAD23) are DNA repair factors participate in the ubiquitin/proteasome system (UPS).  Although the genome-wide analysis of RAD23 family members has been conducted in some species, little is known about RAD23 genes in apple (Malus×domestica Borkh.).  We analyzed this gene family in M. domestica in terms of genomic locations, protein and promoter structures, and expressions in response to stresses.  Various members showed a ubiquitous pattern of expression in all selected apple parts.  Their expressions were altered under chilling, heat, and hydrogen peroxide treatments, as well as abscisic acid (ABA) treatment and water deficiency, suggesting their possible roles in plant stress responses.  These results provide essential information about RAD23 genes in apple and will contribute to further functional studies

Zinc (Zn) deficiency is widespread among citrus plants, but information about the mechanisms for Zn deficiency response in these plants is scarce. In the present study, different navel orange (Citrus sinensis (L.) Osbeck) leaves with various yellowing levels were sampled in our experimental orchard, and upon estimation of nutrient contents, Zn deficiencies were diagnosed as mild, moderate, and severe. Further analysis of chlorophyll content, photosynthetic characteristics, antioxidant enzyme activities, and expression levels of Zn/Iron-regulated transporter-like protein (ZIP) family genes were conducted in the sampled Zn-deficient leaves. The results showed that chlorophyll contents and net photosynthetic rate (P_n) seemed to decrease with reduced Zn contents. In addition, comparison of severe Zn-deficient and normal leaves revealed that activities of peroxidase (POD) and catalase (CAT) increased significantly, whereas that of Zn-containing enzymes such as Cu/Zn superoxide dismutase (Cu/Zn-SOD) significantly reduced with decreasing Zn contents. As expected, expression of the ZIP family genes, ZIP1, ZIP3, and ZIP4, was induced by Zn deficiencies. These results deepen our understanding of Zn deficiency in citrus plants as well as provide useful preliminary information for further research.

China is in a dominant position in apple production globally with both the largest apple growing area and the largest export of fresh apple fruits. However, the annual productivity of China’s apple is significantly lower than that of other dominant apple producing countries. In addition, apple production is based on excessive application of chemical fertilizers and the nutrient use efficiency (especially nitrogen) is therefore low and the nutrient emissions to the environment are high. Apple production in China is considerably contributes to farmers’ incomes and is important as export product. There is an urgent need to enhance apple productivity and improve nutrient use efficiencies in intensive apple production systems in the country. These can be attained by improved understanding of production potential, yield gaps, nutrient use and best management in apple orchards. To the end, priorities in research on apple production systems and required political support are described which may lead to more sustainable and environmental-friendly intensification of apple production in China.

    The vacuolar proton pump ATPase (V-H⁺-ATPase), which is a multi-subunit membrane protein complex, plays a major role in the activation of ion and nutrient transport and has been suggested to be involved in several physiological processes, such as cell expansion and salt tolerance. In this study, three genes encoding V-H⁺-ATPase subunits B (ScVHA-B, GenBank: JF826506), C (ScVHA-C, GenBank: JF826507) and H (ScVHA-H, GenBank: JF826508) were isolated from the halophyte Suaeda corniculata. The transcript levels of ScVHA-B, ScVHA-C and ScVHA-H were increased by salt, drought and saline-alkali treatments. V-H⁺-ATPase activity was also examined under salt, drought and saline-alkali stresses. The results showed that V-H⁺-ATPase activity was correlated with salt, drought and saline-alkali stress. Furthermore, V-H⁺-ATPase subunits B, C and H (ScVHA-B, ScVHA-C and ScVHA-H) from S. corniculata were introduced separately into the alfalfa genome. The transgenic alfalfa was verified by Southern and Northern blot analysis. During salt and saline-alkali stresses, transgenic linevacuolar proton pump, salt tolerance, saline-alkali tolerance, alfalfa
s carrying the B, C and H subunits had higher germination rates than the wild type (WT). More free proline, higher superoxide dismutase (SOD) activity and lower malondialdehyde (MDA) levels were detected in the transgenic plants under salt and saline-alkali treatments. Moreover, the ScVHA-B transgenic lines showed greater tolerance to salt and saline-alkali stresses than the WT. These results suggest that overexpression of ScVHA-B, ScVHA-C and ScVHA-H improves tolerance to salt and saline-alkali stresses in transgenic alfalfa.

Aeromonas hydrophila isolates from clinical cases (n=43) were tested against 8 antimicrobial agents and typed by outer membrane protein (OMP) pattern by using sodium dodecyl sulfate gel electrophoresis. All isolates were resistant to ampicillin (MICs, 16 mg mL-1) and sulfamonomethoxine (MICs, 64 mg mL-1), but susceptible to norfloxacin (MICs, 0.5 mg mL-1). There was a high incidence of resistance to erythromycin (90.70%) and tylosin (93.02%), while a low incidences of resistance to ciprofloxacin (2.33%), enrofloxacin (2.33%) and florfenicol (4.65%). Six different outer membrane protein patterns were found among 34 isolates by analyzing proteins in the range of 22 to 50 kDa, other than 9 isolates with their respective profiles. The strains with the similar OMP profiles had similar resistances. Compared with the other strains from the same OMP patterns, NB-1, A.Pun and MR-1 had lacked the proteins in the range of 30 to 45 kDa and their resistance to florfenicol substantially increased. It is speculated that the outer membrane protein changes might correlate with decreased susceptibility to florfenicol in the three strains. Some strains which showed completely identical OMP types had a little difference in their resistance to fluoroquinolones, indicating that there might be other factors that were involved in the antimicrobial resistance of A. hydrophila.