Development of a method for live octopus (Octopus vulgaris) transportation for long distance at high densities

The development of new octopus‐based products with a growing economic value ensures the commercial interest of this species, making live octopus export an activity of great interest. The aim of this study was to develop a method for long‐distance transportation of live octopus at high densities. The system was composed by 220‐L tanks with cooling and aeration, where the animals were kept separated from each others. The water temperature was maintained at 10°C, after a decreasing of 1°C/hr. Live octopus transportation was tested for 48 hr at two densities: 50 kg/m³ and 100 kg/m³. During this period, water parameters were monitored. Stress response was evaluated through the analysis of haemolymph, muscle and brain tissues. No mortality was registered after 48 hr for both treatments. In all trials, water quality remained within the normal limits in both densities; there was, however, a significative increase of ammonia levels in the water. Ammonia, dopamine and Hsp70 levels were analysed in the beginning and at the end of the experiment for both densities; however, no significant differences were found among them. In general, this system seems to be a viable solution for live octopus 48‐hr transportation at a density of 100 kg/m³.     

云南小麦品种（系）锈病和赤霉病抗性功能基因的KASP标记检测

利用5个锈病成株期抗性基因的KASP标记Sr2_ger9 3p、Lr34jagger、CSTM4_67G、Lr68-2、VPM_SNP和抗赤霉病基因Fhb1的KASP标记TaHRC-KASP,对云南省育成的42个小麦品种（系）进行检测,旨在筛选出含有目标基因的优异小麦种质,为云南省持久抗病小麦新品种（系）的选育提供材料。结果表明,4个材料含兼抗型成株抗锈病基因Lr34/Yr18/Sr57,频率为9.52%;6个品种（系）含兼抗型成株抗锈病基因Lr67/Yr46/Sr55,频率为14.29%;7个材料含抗慢叶锈病基因Lr68,频率为16.67%;含兼抗型成株抗锈病基因Sr2/Yr30和成株抗叶锈基因Lr37的材料各有1个,频率均为2.38%;未检测出含抗赤霉病基因Fhb1的品种（系）。云麦69、云麦75、云麦56、宜麦1号和宜麦3号等兼有2个成株期抗锈病基因,可作为今后云南持久抗锈病育种的抗源材料。     

茄子反向温敏雄性不育系可溶性糖含量及相关基因表达分析

为揭示茄子反向温敏核雄性不育(rTGMS)发生与糖类代谢的关系,以茄子rTGMS系‘05ms’及温度不敏感系‘S63’为试材,对不育期和可育期的叶片及花蕾的可溶性糖含量进行比较,结果表明,‘05ms’在不育和可育时期叶片和花蕾中的可溶性糖含量均显著低于‘S63’;进一步对‘05ms’花蕾进行转录组测序及对其糖类代谢相关途径进行分析,发现差异表达基因主要富集在3个通路:淀粉和蔗糖代谢、糖酵解/糖异生和氨基糖和核苷酸糖代谢,其中淀粉和蔗糖代谢通路的差异基因最多且大部分在低温条件表达下调,主要影响了葡萄糖和半乳糖醛酸的合成。此结果初步揭示了茄子反向温敏雄性不育系中糖类代谢响应温度变化的特点,为茄子雄性不育杂交育种提供理论依据。     

蓖麻GST的基因克隆及生物信息学分析

谷胱甘肽S-转移酶(glutathione S-transferase,GST)是谷胱甘肽结合反应的关键酶,催化谷胱甘肽结合反应的起始步骤,主要存在于胞液中,有多种形式,其在病理学上有一定的重要性。本研究通过克隆GST基因以及对其进行相关的生物信息学分析。利用聚合酶链式反应(polymerase chain reaction,PCR)技术扩增该基因,经分析该基因可编码219个氨基酸,并且通过相关分析得到这219个氨基酸组成的蛋白为稳定的亲水性蛋白,有一个明显的区段存在信号肽,在该蛋白中α-螺旋和无规则卷曲是主要的二级结构方式,根据序列同源比对,蓖麻、麻疯树和榴莲聚类到一起,说明相对于其他物种,它们的同源性较高。这些分析结果为蓖麻谷胱甘肽S-转移酶生物合成及其功能的进一步研究提供一定的理论与实践依据。     

Outer membrane protein A of Acinetobacter baumannii induces autophagy via Akt/mTOR/p70S6K signaling pathway in RAW264.7 cells

AIM:To analyze the effects of outer membrane protein A (OmpA) from Acinetobacter baumannii ATCC 19606 on the autophagy of RAW264.7 cells. METHODS:The RAW264.7 cell model stimulated by OmpA was established. The effects of OmpA on the autophagy of RAW264.7 cells were detected by immunofluorescence, Western blot and transmission electron microscopy. RESULTS:The OmpA increased the expression of LC3B-Ⅱ and reduced the phosphorylation levels of Akt, mTOR and p70S6K. Rapamycin further reduced the phosphorylation levels of mTOR and p-70S6K, and increased the expression of LC3B-Ⅱ induced by OmpA. CONCLUSION:The OmpA of Acinetobacter baumannii induces autophagy via Akt/mTOR/p70S6K signaling pathway in the RAW264.7 cells. This work provides a basis for further research on the molecular mechanism of autophagy induced by Acinetobacter baumannii to find a new method against the infection of Acinetobacter baumannii.     

中国杧果科学研究70年

新中国成立70年来,我国杧果科学研究在人才培养与团队建设、硬件平台、基础理论创新与技术研发等方面均取得了重要进展,有力支撑了我国杧果产业的创建和可持续发展。本文在概述世界和我国杧果地位、分布、规模的基础上,从科学研究发展历程、种质资源与遗传育种、土壤与肥水管理、耕作模式与花果管理、病虫害防控、采后贮运保鲜、产业经济等方面全面梳理了我国杧果研究取得的成就,分析了存在的不足并提出了未来的重点发展方向。     

文心兰HSP70基因的克隆及表达分析

为研究文心兰热激蛋白70基因（命名为OnHSP70）的分子特性及表达特点,以文心兰‘柠檬绿’（Oncidium hybridum ‘Honey Angel’）为材料,采用RT-PCR技术克隆OnHSP70,利用生物信息学方法分析其分子特性,通过qRT-PCR技术分析其在不同组织及不同非生物胁迫处理下的表达特点。生物信息学分析表明：该基因开放阅读框为1944 bp,编码647个氨基酸,翻译的蛋白为稳定蛋白,属于HSP70超家族,其蛋白二级结构由41.11%的α-螺旋、17.77%的延伸链、7.73%的β-转角和33.38%的无规卷曲组成,具有2个高度保守的功能域。聚类分析表明：该蛋白与铁皮石斛和深圳拟兰的HSP70亲缘关系较近。qRT-PCR分析表明：文心兰HSP70在4种不同组织器官中具有表达差异性,在花中的表达量最高;高温处理后基因的表达量明显上升;40 ℃高温胁迫4 h时表达量达峰值;茉莉酸甲酯及水杨酸处理时表达量呈现下调响应。本研究为后期该基因功能研究及提高文心兰对高温的适应性提供理论基础。     

NaCl胁迫对甜瓜生理指标及相关基因表达的影响

以薄皮甜瓜(IVF05、IVF89)和厚皮甜瓜(IVF117、IVF303)为试材,研究Na Cl(150 mmol·L~(-1))处理对4个甜瓜品种发芽、幼苗生长及相关基因表达的影响。结果表明,在Na Cl胁迫下,薄皮甜瓜IVF89的生长指标受到的抑制作用最大,其发芽率、发芽指数、胚根长度和胚轴长度分别下降23百分点、76.24%、46.71%和39.27%;厚皮甜瓜IVF303的生长指标受到的抑制作用最小,其发芽率下降不明显,发芽指数、胚根长度和胚轴长度分别下降46.52%、34.86%和23.08%;Na Cl胁迫提高了甜瓜幼苗叶片中超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)的活性,增加了游离脯氨酸(Pro)、丙二醛(MDA)、可溶性蛋白的含量;4个甜瓜品种幼苗的净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)和SPAD值均显著降低,IVF05、IVF89、IVF117和IVF303的Pn分别下降了64.44%、69.62%、61.23%、60.00%;4个甜瓜品种的卡尔文循环(Calvin)关键酶基因1,5-二磷酸核酮糖羧化酶/加氧酶基因(CmRubisco)、3-磷酸甘油酸激酶基因(CmPGK)和叶绿体型果糖-1,6-二磷酸酶基因(Cm FBPase)表达均显著下调,核酮糖-5-磷酸激酶基因(Cm PRK)表达大多上调;甜瓜耐盐相关基因质膜Na~+/H~+逆向转运蛋白基因(Cm SOS1)、液泡膜Na~+/H~+逆向转运蛋白基因(CmNHX1)、高亲和力K~+转运体基因(CmHKT1)和N-乙酰氨基葡萄糖基转移酶基因(CmGnT)的表达均上调,其中IVF303的基因表达上调幅度最大。通过主成分分析、相关分析和聚类分析等综合分析方法,将4个甜瓜品种分为3类,IVF05和IVF89为盐敏感类型,IVF117为中耐盐类型,IVF303为强耐盐类型。     

大豆RING/U-box蛋白Glyma.13G115900的克隆及其对非生物胁迫的应答

课题组前期对干旱胁迫下大豆转录组的数据分析发现大豆Glyma.13G115900基因编码一个RING/U-box蛋白,其表达水平受干旱胁迫影响显著。本研究以垦丰16大豆为试验材料,克隆Glyma.13G115900基因。氨基酸多重序列比对表明其编码的蛋白与其它物种都具有高度保守的RING/U-box结构域。构建原核表达载体pET-29b-Glyma.13G115900转化到大肠杆菌中,Glyma.13G115900蛋白在大肠杆菌中能够表达。荧光定量PCR分析表明Glyma.13G115900基因的表达量受PEG、NaCl和ABA的影响显著,但基本不受冷胁迫的诱导。经PEG和NaCl处理后,该基因表达量与CK相比呈现出显著下降的趋势,PEG处理的表达量变化比NaCl下调的更明显;在ABA诱导下与CK相比该基因的mRNA丰度呈现出先上升后下降的趋势,在4 h表达量出现峰值,推测该基因可能通过依赖于ABA途径参与非生物胁迫应答。以上结果为进一步深入研究该基因的调控途径奠定基础。     

Responses of glutamine synthetase activity and gene expression to nitrogen levels in winter wheat cultivars with different grain protein content

Glutamine synthetase (GS) plays a central role in plant nitrogen (N) metabolism, which improves crops grain protein content. A pot experiment in field condition was carried out to evaluate GS expression and activity, and grain protein content in high (Wanmai16) and low grain protein (Loumai24) wheat cultivars under two N levels (0.05 and 0.15 g N kg⁻¹ soil). High nitrogen (HN) resulted in significant increases in GS1 and GS2 expression at 10 days after anthesis (DAA), and higher GS activity during the entire grain filling stage. HN also significantly increased yield, grain protein content and protein fraction (except for glutenin of Luomai24) in two wheat cultivars, which indicated that it increased grain yield and protein content by improving nitrogen metabolism. Wanmai16 showed higher grain protein content, gliadin and glutenin content, and had higher expression level of GS2 both in flag leaves and grains at early grain filling stage. However, Luomai24 had greater yield and higher expression level of GS1. The difference expression of GS2 and GS1 genes indicates they had various contributions to the accumulation of protein and starch in wheat grains, respectively. The results suggest that GS2 would be serving as a potential breeding target for improving wheat quality.