Hippophae rhamnoides L. is extensively distributed throughout China and plays a pioneering role in combating desertification and soil erosion in northern regions. H. rhamnoides contains abundant nutrients and is of medical and economic value. However, there has been a lack of research on sea buckthorn seeds, both domestically and internationally, particularly regarding the mechanisms governing their growth and germination. Therefore, to explore the growth of sea buckthorn seeds, this study analyzed and studied the molecular mechanism of seed germination process of sea buckthorn. To better understand the molecular mechanism underlying seed germination in sea buckthorn, we used transcriptomics to compare gene expression before and after seed germination in H. rhamnoides subsp. sinensis Rousi. We identified 9,988 differentially expressed genes (5,593 upregulated and 4,395 downregulated). A bioinformatics-based analysis revealed that changes in plant hormone signal transduction and starch and sucrose metabolism-related gene expression may regulate seed germination in this species. Notably, the levels of auxin (IAA), cytokinin (CTK), and brassinosteroids (BR) increased during seed germination while those of ABA decreased. Exogenous application of IAA, CTK, and BR promoted sea buckthorn seed germination, while ABA inhibited it. These findings suggested that hormones play an important role in the process of sea buckthorn seed germination. This study provides preliminary information about the seed germination mechanism in sea buckthorn, offering an essential reference for improving seed breeding and germplasm and laying the foundation for further resistance research on the molecular mechanism of seed germination of sea buckthorn in this species.

蜡质诱导因子 1(wax inducer 1/SHN1,WIN1/SHN1)与植物的抗逆性密切相关.为研究WIN1 在耐旱经济灌木扁果枸杞(Lycium barbarum ssp.Bianguo)中的功能及其应用前景,本研究用反转录聚合酶链式反应(RT-PCR)技术从扁果枸杞中克隆了LbWIN1,构建了LbWIN1 融合增强型绿色荧光蛋白(EGFP)的载体并表达在拟南芥(Arabidopsis thaliana)原生质体中进行亚细胞定位,用实时荧光定量PCR 法(quantitative real-time PCR,qRT-PCR)分析 LbWIN1 的表达特征.结果表明,LbWIN1 基因长 1 103 bp,含 600 bp的开放阅读框,编码 199 个氨基酸,含有 1 个高度保守的 AP2 结构域.系统进化树分析发现,LbWIN1 蛋白与茄科番茄(Solanum lycopersicum)SlWIN1 和辣椒(Capsicum chinense)CcWIN1 的同源性高达 99%.亚细胞定位分析证明LbWIN1 蛋白定位于细胞核.LbWIN1 基因在叶、茎、根中均表达,其表达量受NaCl、渗透胁迫和外源脱落酸(ABA)诱导,表明LbWIN1 参与旱生植物响应盐和干旱等非生物胁迫.

细胞增殖调控是细胞生物学教学的重点和难点内容,精巧的教学设计有助于提高学生学习兴趣和效果.本文分析了细胞增殖调控部分的教学内容,设计了基于问题驱动,结合思维导图的教学过程.本教学设计能通过变换问题顺序、删减问题等调整授课内容,达到优化教学重点的目的.本教学设计逻辑严密,通过层层推进的问题展开教学内容,并且注重学生主动参与教学,设计了大量教学互动环节.本教学设计有助于提高学生逻辑思维能力、实验设计能力和协作能力.

The perennial shrub of Calligonum mongolicum Turcz. is a dominant native plant in all Calligonum species, which has the largest and most widespread geographic distribution in arid deserts of northern China. Understanding the phylogenetic relationship between C. mongolicum and closely related plant species will offer guidance on the classification and identification of inter-species and their varieties. The chloroplast (cp) genome is an optimal model to decipher phylogenetic relationships in related plant families. In the present study, the complete cp genome of C. mongolicum was sequenced and compared with other Calligonum species. The cp genome of C. mongolicum was 162,124 bp in length with a quadripartite structure. A total of 131 functional genes were annotated, 14 different genes of which harbored introns and exons, 50 long repeat sequences and 244 simple sequences repeats were identified. Synonymous codon usage (SCU) analysis of C. mongolicum and other 8 Calligonum species exhibited similar change patterns, and natural selection would be crucial for SCU bias in all the 9 Calligonum species. The phylogenetic tree showed that C. mongolicum was clustered closely to Calligonum species. From the comparative analysis of genomic structures, the inverted repeat regions (IRa and IRb) were less divergent than other parts and coding regions was relatively conserved than non-coding regions. Compared to other Calligonum species, the IR region of C. mongolicum had a slight contraction. Our results comprehensively depict the architecture of C. mongolicum cp genome, and will lay a vigorous foundation for further study on molecular marker selection, phylogenetic analysis, and population researches in Calligonum species.

The succulent xerophyte Zygophyllum xanthoxylum (Bunge) Engl. can absorb Na+ from the soil as an osmoticum in order to resist osmotic stress. The tonoplast Na+/H+ antiporter ZxNHX1 is essential for maintaining the salt-accumulation characteristics of Z. xanthoxylum by compartmentalizing Na+ into vacuoles. Previous results revealed that the silencing of ZxNHX1 greatly decreased Na+ accumulation in Z. xanthoxylum under 50 mM NaCl due to the weakened compartmentalisation; in addition, K+ concentration also significantly reduced in ZxNHX1-RNAi lines. Yet, whether the reduction of K+ concentration was directly triggered by the silencing of ZxNHX1 remains unclear. In this study, the growth parameters and expression levels of ZxSOS1, ZxHKT1;1, ZxAKT1 and ZxSKOR were measured in wild-type and ZxNHX1-RNAi lines under control or -0.5 MPa osmotic stress. The results showed that the silencing of ZxNHX1 inhibited the plant growth, decreased Na+ concentration in leaves, reduced the transcript abundance of ZxSOS1 and dramatically increased that of ZxHKT1;1 in roots of Z. xanthoxylum under osmotic stress; whereas tissue K+ concentrations and the expression level of ZxSKOR displayed no significant variations, and the expression of ZxAKT1 were significantly reduced in ZxNHX1-RNAi lines under osmotic stress, compared with the wild type. These results suggest that in Z. xanthoxylum, ZxNHX1 can maintain the normal growth by compartmentalizing Na+ into vacuoles, and regulate the spatial distribution of Na+ indirectly by affecting the expressions of ZxSOS1 and ZxHKT1;1. Moreover, the silencing of ZxNHX1 is not the main reason that led to the reduction of K+ concentration in ZxNHX1-RNAi lines under 50 mM NaCl, and ZxNHX1 might be indirectly involved in regulating K+ homeostasis.

Cuticular waxes, a waterproofing hydrophobic barrier that coats the aerial surfaces of all terrestrial plants, protects plants from various environmental stresses, particularly drought stress. The molecular basis of cuticular wax deposition in xerophytes (e.g. desert plants), including transport mechanisms of waxes to the cell wall, is poorly understood. Here, ZxABCG11 encoding an ATP binding cassette (ABC) transporter was isolated from the xerophyte Zygophyllum xanthoxylum, since we found it was predominantly expressed in young leaves and induced by salt, drought, heat, and combined drought and heat stresses. To ascertain the function of ZxABCG11, we introduced ZxABCG11 into Arabidopsis thaliana and found an increased abundance of cuticular wax on stems and leaves of the transgenic plants, which also exhibited a thickened cuticle and reduced cuticle permeability. Transcript expression analysis revealed that many genes related to wax biosynthesis and transport were up-regulated in these ZxABCG11 transgenic plants. Furthermore, compared with wild type Arabidopsis, ZxABCG11 transgenic plants displayed a higher biomass and survival rate under drought conditions. Taken together, we propose that the expression of ZxABCG11 in Arabidopsis promotes cuticular wax accumulation and thicker cuticle formation, thereby enhancing drought tolerance.

为鉴定醛脱羰基酶(ECERIFERUM1,CER1)基因在旱生植物表皮蜡质烷烃合成过程中的功能,培育适宜干制加工品种,选定扁果枸杞(Lycium barbarum ssp.Bianguo)LbCER1基因1469～1840 bp区段为最佳RNA干扰(RNA interference,RNAi)靶序列,克隆该区段并将其插入pKANNIBAL载体,构建具有发卡状反向重复序列的RNAi中间载体pKANNIBAL-LbCER1(+)-PDK intron-LbCER1(-),并将发卡状反向重复序列转入RNAi表达载体pART27,再用冻融法将pART27-LbCER1(+)-PDK intron-LbCER1(-)载体转化根癌农杆菌(Agrobacterium tumefa-ciens)GV3101,并用限制性内切酶双酶切法和PCR法检测转化的农杆菌菌株.结果表明,RNAi的中间载体和表达载体均结构正确,并且RNAi表达载体pART27-LbCER1(+)-PDK intron-LbCER1(-)成功转化根癌农杆菌GV3101,说明该构建方法正确,将为进一步筛选LbCER1-RNAi植株,获得适宜干制加工的宁夏枸杞新品种奠定基础.

BackgroundThe perennial shrub of Calligonum mongolicum is a dominant native plant in all Calligonum species, which has the largest and most widespread geographic distribution in arid deserts of northern China. Understanding the phylogenetic relationship between C. mongolicum and closely related plant species will offer guidance on the classification and identification of inter-species and their varieties. The chloroplast (cp) genome is an optimal model to decipher phylogenetic relationships and genome evolution in related plant families. In the present study, the complete cp genome of C. mongolicum was sequenced, and the characteristics were described, then the genomic structure was compared to other three Polygonaceae species.ResultsThe cp genome of C. mongolicum was 162,124 bp in length with a quadripartite structure. A total of 131 functional genes were annotated, 14 different genes of which harbored introns and exons, 50 long repeat sequences and 244 simple sequences repeats were identified. Synonymous codon usage (SCU) analysis exhibited A/T preference, and 7 codons were identified as the optimal codons. Multivariate statistical analysis of parity rule 2, ENC-plot, and neutrality plot were combined conducted to imply natural selection as the crucial constraint in SCU bias in C. mongolicum cp genome.The phylogenetic tree showed that Rumex acetosa was the most related plant to C. mongolicum . From the comparative analysis of genomic structures, the inverted repeat regions (IRa and IRb) were less divergent than other parts and coding regions was relatively conserved than non-coding regions. Compared to other species in the Polygonaceae, the borders of IRb/SSC and SSC/IRa in C. mongolicum changed greatly. Furthermore, adaptive evolution analysis of 75 orthologous protein-coding genes indicated that only the psbK gene was under positive selection, which might be crucial in the adaptive evolution of C. mongolicum .ConclusionsOur results comprehensively depicts the architecture of C. mongolicum cp genome, and will lay a vigorous foundation for further study on molecular marker selection, phylogenetic analysis, and population researches in Calligonum species.

在单因素试验的基础上,通过正交试验法优化苦水玫瑰花红色素的提取条件,并对其稳定性进行研究.结果 表明,苦水玫瑰花红色素的最佳提取条件为:提取剂选用80％的乙醇,pH为2,料液比1∶60(g/mL),浸提时间60 min;苦水玫瑰花红色素在pH＜6的酸性条件下较稳定,在碱性介质中极不稳定,在70℃以下较稳定,但对光较敏感,食品添加剂蔗糖和VC对其稳定性影响不大(色素吸光值变化幅度在10％以内).该色素对还原剂Na2S2O3有一定的耐受能力,而对氧化剂H2O2的耐受性较弱,金属离子Na+、Al3+、K+、Ca2+对色素的稳定性无明显影响,而Fe2+和Fe3+对该色素的稳定性影响较大.

Flavonoids in astragali radix exert biological health functions actions such as antioxidant and immune-enhancing effects. They are often added to food and feed as immunopotentiators. The effects of ethanol concentration, the solid/liquid ratio, and extraction time on the extraction content of total flavonoids (TFA) from Astragalus membranaceus were evaluated using single factor experiments. Furthermore, the optimum beneficiation and depuration technology of TFA was investigated with HPD722 macroporous resin-polyamide combination method. The results showed that the extraction content of TFA and relative standard deviation (RSD) were 2.78 mg·g–1 and 1.41%, respectively, under the conditions of 70% ethanol concentration, solid/liquid ratio of 1∶17, and an extraction time of 70 min. When TFA were enriched and purified using HPD722 macroporous resin-polyamide, the extraction content of TFA was 311 mg·g–1 and its purity was increased 112 times. As the main component of TFA, the content of calycosin was increased 327 times from 0.13 mg·g–1 to 42.56 mg·g–1 and formononetin was increased 132 times from 0.06 mg·g–1 to 7.89 mg·g–1.

为克隆扁果枸杞(Lycium barbarum Bianguo)肌动蛋白基因作为基因表达模式分析的内参基因,根据几种茄科植物肌动蛋白氨基酸序列保守区设计引物,以扁果枸杞3周龄叶总RNA为模板,采用RT-PCR的方法扩增肌动蛋白基因片段并连接到pMD18-T载体上,阳性克隆经PCR检测后测序.结果表明,该基因片段长598 bp,编码198个氨基酸,与枸杞(Lycium chinense)核苷酸序列的相似度和同源性分别达97％和100％,说明是肌动蛋白基因片段,命名为LbACT.荧光定量PCR分析表明,盐处理下LbACT基因在各器官中的Ct值稳定,可作为内参基因用于研究扁果构杞功能基因的表达模式分析.

为研究旱生植物中醛脱羰基酶基因(ECERIFERUM1,CER1)在响应渗透胁迫过程中的分子特性,用逆转录PCR(Reverse transcription PCR,RT-PCR)和RACE法克隆了耐盐耐旱的扁果枸杞CER1基因的cDNA序列,对该序列进行了生物信息学分析,并用实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)法检测在渗透胁迫处理后扁果枸杞CER1在各器官中的转录水平.结果表明,扁果枸杞CER1的cDNA序列长2168 bp,开放阅读框为1881 bp,命名为LbCER1,编码626个氨基酸,有4个跨膜区、1个脂肪酸羟化酶结构域和1个位于C-末端的Wax2结构域.该蛋白的理论分子质量为72.47 ku,等电点为7.40,脂肪指数为91.28,不稳定系数为27.88,平均亲水性为-0.070,是一种耐高温、性质稳定的偏碱性亲水蛋白.LbCER1蛋白定位在内质网膜上,二级结构主要由α-螺旋和环组成,分别占44.09％和45.21％,与三级结构预测结果相符.LbCER1与茄科植物亲缘关系近,同源性达100％.LbCER1基因在叶、茎、根中均表达.与对照相比,160 mmol/L山梨醇渗透胁迫处理下,6 h时LbCER1基因在叶中的相对表达量最高,增加1.8倍,茎中的相对表达量增加21％,12 h时降低14％,根中渗透胁迫处理6,12 h时,LbCER1的相对表达量分别增加1.1倍和32％;80 mmol/L山梨醇渗透胁迫下,仅叶在处理6 h时LbCER1表达量增加66％,茎和根中表达量降低或差异不显著,表明LbCER1可能与扁果枸杞响应渗透胁迫有关.

实践教学体系是高等学校实现工程类专业人才培养目标的重要保证.在多年的实践探索中,兰州理工大学生物工程专业构建了以实验教学、实习实训、课程设计和创新创业等为主要内容的实践教学体系,建立了较为完备的校内实验室以及稳定的校外实习实训基地,为该专业创新型人才培养搭建了坚实的平台.

为探讨构建不同品种宁夏枸杞鲜果适宜干制加工的鲜果品质性状综合评价体系,以西北地区广泛栽培的9个品种宁夏枸杞鲜果为材料,用扫描电镜研究了表皮蜡质形态,并通过单因素方差分析、因子分析、相关性分析和聚类分析,比较了比表面积、组织含水量、单果体积、百粒鲜重和百粒干重的差异.结果表明,9个品种宁夏枸杞鲜果果形和表皮蜡质形态相似,但品种间比表面积、组织含水量、单果体积、百粒鲜重和百粒干重差异显著.组织含水量与比表面积呈显著正相关,与百粒干重呈极显著负相关;体积、百粒鲜重、百粒干重间均呈极显著正相关.百粒干重和组织含水量是体现枸杞鲜果干制加工特性的关键指标.9个宁夏枸杞品种聚为4类,蒙杞1号为一类,最适宜于干制;宁杞0901、宁杞7号、宁杞3号聚为一类;宁杞0702号、宁杞6号、宁杞5号、宁杞4号聚为一类,扁果枸杞单独聚为一类,后两类不宜干制.

Aim To studv the chemical eonstituents of the flower buds of Tussilago farfara L.in the China National GAP Base of Traditional Chinese Materia Medica and provide scientific basis for quality control. Methods The constituents were separated and purified by different chromatographic methods,and their structures were elucidated by IR.MS and NMR techniques.Results Twenty eight compounds were isolated from the flower buds of T.farfara.Their structures were identified as nheptacosane (1),bis(2-ethylhexyl)phthalate (2),7β-[3'-ethylcrotonoyloxy]-la-[2'-methylbutyryloxy]-3,14-dehydro-Z-notonipetranone (3),7β-[3'-ethylcrotonoyloxy]-la-[2'-methylbutyryloxy]-3,14-dehydro-E-notonipetranone(4),tussilagone(5),dibutyl phthatate dihydroxypropylpalmitate(16),daucosterol(17),6-hydroxy-2,6-dimethylhept-2-en-4-one(18),femlic acid(19),isofemlic acid (27),and adenosine (28).Conclusion Compounds 1,12-16,18 and 20 were obtained from the genus Tussilago for the first time.

Potassium (K+) is one of the major nutrients,essential for plant growth and development.Inward rectifying K+ channel AKT1 belongs to Shaker-type K+ channels,and a K+ sensor in the plasma membrane has been shown to play an important role in mediating K+ uptake.AKT1 is also essential for normal growth and development,stomatal action and transpiration,and improving the drought and salt tolerance of plants.In this review,we consider recent advances in molecular structure,localization,expression,regulation and function of AKT1.Finally,research directions for the future are proposed,including the use of proteomics,gene engineering technology,and posttranscriptional gene silencing,in order to further investigate the role of AKT1 in plant physiological metabolism.

马铃薯块茎低温糖化现象除了直接受到淀粉和糖代谢相关酶如淀粉酶、酸性转化酶的活性影响外,也可能间接受到赤霉素和脱落酸等植物激素的调控.以马铃薯品种'大西洋'块茎为试验材料,采用50 mmoL GA3处理12 h后置于低温((4±0.5)℃)和室温((22±1)℃)条件下贮藏10 d,测定薯块的炸片色泽、还原糖含量、淀粉酶和酸性转化酶活性,研究赤霉素对马铃薯块茎低温糖化的影响.结果表明,GA3处理使马铃薯块茎的炸片色泽加深,还原糖含量增加121.1%;同时,GA3处理提高了淀粉酶的活性,而对酸性转化酶的活性没有影响.由此表明,GA3可能通过提高马铃薯块茎淀粉酶的活性而使还原糖积累,从而导致炸片色泽加深即增强了低温糖化现象.

High concentration of sucrose can significantly promot potato tubeization.Sucrose can not only maintain the growth of potato plant,but also specifically regulate the expression of tuberization-related genes. Moreover,sucrose is also involved in the process of regulating tuberization by hormones. This paper reviews the research progress made in studying mechanism of regulating potato tuberization by sucrose.