辣椒高赖氨酸蛋白基因Cflr全长cDNA的克隆及其组织表达特征

为了在茄科植物中寻找新的高赖氨酸蛋白基因,以辣椒栽培种‘江蔬7号'的成熟花粉为材料,根据马铃薯和番茄中高赖氨酸蛋白基因的保守序列设计引物,通过RT-PCR技术扩增到一条长390 bp的cDNA片段,继而应用RACE技术克隆了一个辣椒高赖氨酸蛋白基因的全长cDNA,命名为Cflr（GenBank登录号：EU367999）。Cflr基因cDNA全长920 bp,5＇端有84 bp的非翻译区,3＇端具有完整的polyA尾,包含一个编码223个氨基酸的完整开放阅读框。Cflr基因与马铃薯和番茄高赖氨酸蛋白基因cDNA序列的同源性为50%～60%,氨基酸序列的同源性为40%～50%。该基因所编码的蛋白质中赖氨酸含量为21.2%,苏氨酸含量为10.3%,是目前已知赖氨酸含量最高的天然蛋白。半定量RT-PCR结果表明,Cflr基因在辣椒未成熟花药、成熟花粉、花瓣、茎、叶片和根中均有表达,在成熟花粉和花瓣中的表达丰度最高,在叶片中表达量明显减少,而在未成熟花药、茎和根中的表达量极少。     

桦褐孔菌纤维素酶基因的克隆与原核表达

成功获得了桦褐孔菌(Inonotus obliquus)JL 01菌株的内切葡聚糖酶(IO-EG)和β-葡萄糖苷酶(IO-BGL)的cDNA全长序列.eg2基因cDNA序列全长(ORF)为1149 bp,编码382个氨基酸,bgl2基因cDNA序列全长(ORF)为2583 bp,编码860个氨基酸;成功构建了30a-eg2和30a-bgl2大肠杆菌表达菌株,诱导培养后,SDS-PAGE电泳分析表明两个菌株均表达蛋白,且蛋白以包涵体形式存在,为桦褐孔菌高效外源基因表达系统的建立提供了基础.     

八棱海棠肌动蛋白基因(MrACT)的克隆及分析

以八棱海棠(Malus micromalus Makino)为材料,抽取RNA,反转录成cDNA。采用cDNA末端快速扩增(RACE)方法,5’-RACE获得长度1121bp的片段、3’-RACE获得883bp的片段,再通过设计引物扩增得到八棱海棠中一种肌动蛋白基因的cDNA的全长序列。该序列全长1134bp,推测其编码377个氨基酸,等电点为5.16,其编码的氨基酸与拟南芥(Arabidopsis thaliana)actin7编码的氨基酸只有2个氨基酸差别。通过对八棱海棠MrACT基因的表达分析,发现在不同组织中,该基因的表达水平没有明显差异,本文为进一步利用RT-PCR技术,以MrACT基因为内标,研究八棱海棠其他基因的丰度打好基础。     

Pt-CaM基因克隆及对菌核侧耳耐热性影响

通过保守序列比对、cDNA末端快速扩增技术和融合引物巢式PCR技术克隆菌核侧耳（Pleurotus tuber-regium）钙调蛋白（calmodulin,CaM）基因Pt-CaM。基因全长856bp,cDNA全长477bp,编码158个氨基酸。构建Pt-CaM的过表达和干涉载体,利用根癌农杆菌转化技术成功转化至菌核侧耳菌丝体中。在38℃高温培养下,过表达转化子的菌丝较野生型更致密,热激蛋白（heat shock protein）HSP70的表达量为野生型的3.89倍,超氧化物歧化酶（superoxide dismutase,SOD）和过氧化氢酶（catalase,CAT）的活性为野生型的2.49倍和1.83倍;与野生型相比,干涉转化子菌丝生长速率降低,热胁迫损伤加重,其HSP70的表达量为野生型的0.77,SOD和CAT的活性无差别,过氧化物酶（peroxidase,POD）的活性为野生型的0.7。结果表明Pt-CaM可能通过影响HSP70的表达量和SOD、CAT和POD的活性参与菌核侧耳菌丝的耐热性调控。     

龙眼14-3-3基因的克隆与原核表达

 以龙眼(Dimocarpus longan Lour. ) 为试材, 应用同源克隆和RACE方法从花芽中获得了调控 蛋白14-3-3的全长cDNA序列, GenBank登录号为FJ479618 (GI: 218202931) 。该cDNA全长1 121 bp, 包括一个783 bp的开放阅读框, 编码261个氨基酸, 序列比较分析显示14-3-3 cDNA具有较高的保守性。半定量RT-PCR分析结果表明, 14-3-3 mRNA在龙眼叶芽、叶片、花芽和成熟的花中都有表达, 但在花芽中表达量最大。构建了pET14-3-3原核表达系统, 将14-3-3全长cDNA在大肠杆菌中表达, 获得一个分子量约为34 kD的可溶性融合蛋白, 经Western blotting验证, 该蛋白为14-3-3蛋白。     

津田芜菁BrPIP1的克隆及其在非生物胁迫下的表达分析

克隆得到津田芜菁(Brassica rapa ssp.rapifera‘Tsuda’)质膜内在蛋白(plasma membrane intrinsic proteins,PIPs)基因全长cDNA序列,命名为BrPIP1(GenBank登录号为KJ173685),全长为1 056 bp,开放阅读框为861 bp,编码286个氨基酸。荧光定量PCR分析BrPIP1在不同组织以及其在温度、脱水、渗透、ABA和盐等非生物胁迫条件下幼苗中的表达表明,该基因表达具有组织特异性,在花瓣中表达量最高,花蕾中次之;在上述非生物胁迫下表达量都有不同程度增加,暗示BrPIP1在非生物胁迫应答中发挥作用。     

光皮桦BBX类基因BlCOL13的克隆与表达分析

利用RACE、RT-PCR以及基因组步移技术克隆得到光皮桦BlCOL13全长cDNA（GenBank登录号：KP663425）、全长DNA及其启动子序列。该基因DNA全长2 808 bp,具有4个外显子,3个内含子;cDNA全长1 370 bp,包含1 140 bp的开放阅读框,编码含379个氨基酸的蛋白。BlCOL13蛋白N、C端分别有两个B-box和一个CCT结构域,属于典型的BBX蛋白。以拟南芥中BBX蛋白的分类为参照,通过聚类分析表明BlCOL13属于Ⅱ型BBX蛋白;克隆得到的BlCOL13启动子序列长685 bp,含有多个与成花、光响应、激素信号转导以及逆境胁迫相关的顺式作用元件。亚细胞定位试验结果表明该基因主要在细胞核中表达。不同组织中该基因都有一定程度的表达,但在雄花中表达量最高。低温、干旱、ABA和盐胁迫等非生物逆境都在一定程度上诱导BlCOL13的表达,盐胁迫的影响最为明显,表明该基因在光皮桦逆境响应中可能发挥比较重要的作用。     

辣椒高赖氨酸蛋白基因Cflr全长cDNA的克隆及其组织表达特征

为了在茄科植物中寻找新的高赖氨酸蛋白基因,以辣椒栽培种‘江蔬7号'的成熟花粉为材料,根据马铃薯和番茄中高赖氨酸蛋白基因的保守序列设计引物,通过RT-PCR技术扩增到一条长390 bp的cDNA片段,继而应用RACE技术克隆了一个辣椒高赖氨酸蛋白基因的全长cDNA,命名为Cflr（GenBank登录号：EU367999）。Cflr基因cDNA全长920 bp,5＇端有84 bp的非翻译区,3＇端具有完整的polyA尾,包含一个编码223个氨基酸的完整开放阅读框。Cflr基因与马铃薯和番茄高赖氨酸蛋白基因cDNA序列的同源性为50%～60%,氨基酸序列的同源性为40%～50%。该基因所编码的蛋白质中赖氨酸含量为21.2%,苏氨酸含量为10.3%,是目前已知赖氨酸含量最高的天然蛋白。半定量RT-PCR结果表明,Cflr基因在辣椒未成熟花药、成熟花粉、花瓣、茎、叶片和根中均有表达,在成熟花粉和花瓣中的表达丰度最高,在叶片中表达量明显减少,而在未成熟花药、茎和根中的表达量极少。     

Fruit growth,cuticle deposition,water uptake,and fruit cracking in jostaberry,gooseberry, and black currant

Fruit cracking after rain limits the production of a number of crops, including some Ribes species. To gain a better understanding of the factors involved in cracking, fruit growth, deposition of the cuticular membrane (CM), water uptake and fruit cracking were studied in black currant (Ribes nigrum L. cv. Zema), gooseberry (Ribes uva-crispa L. cv. Rote Triumph), and jostaberry (Ribes nidigrolaria B. cv. Jostine). Fruit surface area and fresh mass increased continuously throughout development, whereas deposition of the CM was biphasic. CM mass per fruit increased rapidly up to 42, 41, and 49 days after full bloom (DAFB) in black currant, gooseberry, and jostaberry, respectively. Thereafter, CM mass per fruit remained constant in gooseberry and jostaberry or increased at a lower rate in black currant. The cessation of or reduced rate of CM deposition resulted in a decrease in CM mass per unit area in all berries. Elastic strain of the CM at maturity averaged 23.8% and 19.5% in gooseberry and jostaberry, respectively, and only 8.2% in black currant. Microcracks in the CM were observed first in gooseberry and jostaberry 64 DAFB, whereas there were no microcracks in black currant. Water uptake into mature detached berries was linear over 2 h of incubation. Rates of uptake were highest in gooseberry followed by black currant and jostaberry. Relative uptake was similar via the cut end of the pedicel (32.1%), the apex of the fruit (34.7%) and the fruit surface (33.2%). Rates of water uptake through the fruit surface were positively related to surface area. Average fruit water potential for black currant, gooseberry, and jostaberry was −2.14 ± 0.17, −1.24 ± 0.03, and −1.89 ± 0.20 MPa, while the permeability for osmotic water uptake was 7.7 ± 0.4 × 10⁻⁸, 5.2 ± 0.1 × 10⁻⁸, and 3.3 ± 0.3 × 10⁻⁸ m s⁻¹. Incubating whole fruit in deionized water for 72 h resulted in more cracked jostaberries (94%) than black currants (74%) or gooseberries (50%). A comparison of our findings in Ribes berries with published data for the sweet cherry drupe revealed that the berries fitted the relationships established in sweet cherry among fruit growth, cuticle deposition, strain of the cuticle, microcracking, permeability for osmotic water uptake, frequency of stomata and cracking. The Ribes berries were less susceptible to cracking than sweet cherry.     

Similitude,similarity, and scaling

An exceptionally rich and colorful literature, drawn in almost equal parts from pure mathematics, from the sciences, and from the technologies, has grown up over the years, which bear in different ways on the topics under discussion. It is the intent of the present paper to survey this far-flung literature, point out some of the commonalities and interrelationships which underlie it, and briefly indicate how it has been and can be applied. To my knowledge, this kind of review has not been attempted before.     

Climatic, landform, microtopographic, and overstory canopy controls of tree invasion in a subalpine meadow landscape, Oregon Cascades, USA

Tree invasions have been documented throughout Northern Hemisphere high elevation meadows, as well as globally in many grass and forb-dominated ecosystems. Tree invasions are often associated with large-scale changes in climate or disturbance regimes, but are fundamentally driven by regeneration processes influenced by interactions between climatic, topographic, and biotic factors at multiple spatial scales. The purpose of this research was to quantify spatiotemporal patterns of meadow invasion; and how climate, larger landforms, topography, and overstory trees have interactively influenced tree invasion. We combined airborne Light Detection and Ranging (LiDAR) characterizations of landforms, topography, and overstory vegetation with historical climate, field measurements of snow depth, tree abundance, and tree ages to reconstruct spatial and temporal patterns of tree invasion over five decades in a subalpine meadow complex in the Oregon Cascade Range, USA. Proportion of meadow occupied by trees increased from 8?% in 1950 to 35?% in 2007. Larger landforms, topography, and tree canopies interactively mediated regional climatic controls of tree invasion by modifying depth and persistence of snow pack, while tree canopies also influenced seed source availability. Landscape context played an important role mediating snow depth and tree invasion; on glacial landforms tree invasion was negatively associated with spring snowfall, but on debris flows tree invasion was not associated with snow fall. The importance of snow, uncertain climate change impacts on snow, and mediation of snow by interacting and context dependent factors in complex mountain terrain poses substantial hurdles for understanding how these ecotones may respond to future climate conditions.     

Wetlands, carbon, and climate change

Wetland ecosystems provide an optimum natural environment for the sequestration and long-term storage of carbon dioxide (CO₂) from the atmosphere, yet are natural sources of greenhouse gases emissions, especially methane. We illustrate that most wetlands, when carbon sequestration is compared to methane emissions, do not have 25 times more CO₂ sequestration than methane emissions; therefore, to many landscape managers and non specialists, most wetlands would be considered by some to be sources of climate warming or net radiative forcing. We show by dynamic modeling of carbon flux results from seven detailed studies by us of temperate and tropical wetlands and from 14 other wetland studies by others that methane emissions become unimportant within 300 years compared to carbon sequestration in wetlands. Within that time frame or less, most wetlands become both net carbon and radiative sinks. Furthermore, we estimate that the world’s wetlands, despite being only about 5–8 % of the terrestrial landscape, may currently be net carbon sinks of about 830 Tg/year of carbon with an average of 118 g-C m^?2 year^?1 of net carbon retention. Most of that carbon retention occurs in tropical/subtropical wetlands. We demonstrate that almost all wetlands are net radiative sinks when balancing carbon sequestration and methane emissions and conclude that wetlands can be created and restored to provide C sequestration and other ecosystem services without great concern of creating net radiative sources on the climate due to methane emissions.     

Up-regulation of mRNA expressions of fas,bcl-2, bim,bax, caspase-3, caspase-9, and bcl2l12 in K562 treated with bortezomib

AIM: To detect the treatment of K562 leukemia cells with bortezomib altering the expression of genes fas, bcl-2, bcl2l12, bim, bax, caspase-9 and caspase-3.METHODS: MTT assay was used to detect the inhibition of proliferation. Apoptosis was detected by Annexin-V staining and mitochondrial transmembrane potential (Δψm). RT-PCR was used to analyze the mRNA expressions of fas, bcl-2, bcl2l12, bim, bax, caspase-3 and caspase-9.RESULTS: Bortezomib caused a time- and dose-dependent inhibition of cell proliferation and IC50 of 24 h and 48 h were 161.41 nmol/L and 96.33 nmol/L, respectively. At the concentration of 104 nmol/L, bortezomib induced apoptosis in a time-dependent manner, including increasing annexin-V positivity and decreasing the Δψm. RT-PCR showed that bortezomib up-regulated the mRNA expression of fas, bcl2l12, caspase-9 and caspase-3, but mRNA expressions of bcl-2, bim and bax did not changed obviously.CONCLUSION: Bortezomib inhibits the proliferation of K562 and induces apoptosis, in which fas, bcl2l12, caspase-9 or caspase-3 gene is one of the main genes taking part in.     

氮磷钾硼肥配施对青花菜养分吸收分配及产量和品质的影响

研究了不施肥处理,氮磷钾配施和增施硼肥对青花菜养分吸收分配及产量和品质的影响。结果表明,现蕾期至花球膨大期,青花菜对氮、磷、钾的吸收量最大,整个生长期对钾的吸收最多,氮次之,磷最少。氮磷钾主要分配在叶片中,现蕾后逐渐向花球转移,从现蕾到采收时,叶片中氮、磷、钾分配率分别降低了19.6%、10%和9.0%,花球中氮磷钾分配率分别提高了23.1%、15.9%和11.1%。不施氮、磷、钾肥显著降低了花球产量、维生素C含量和成品率,增施硼肥处理的花球成品率较氮磷钾肥处理提高了4.8%,不施氮肥和钾硫肥(K₂SO₄)显著降低了花球中的硫代葡萄糖苷含量。相关性分析表明,青花菜产量与植株内氮、钾积累量显著正相关,成品率与植株内磷营养显著正相关。     

Principles for forcing tulips,hyacinths, daffodils,Easter lilies and Dutch irises

Systematic investigations on the forcing of tulips, hyacinths, daffodils, Easter lilies and Dutch iris have produced a fairly complete set of scientific principles. In this review, these principles have been classified into a 3-phase concept of forcing. These are: production, programming, and greenhouse. The production phase is defined as all processes which occur during bulb production and it terminates with the harvesting of the bulbs. The programming phase comprises all handling of the bulbs from harvesting until they are placed under greenhouse conditions. The greenhouse phase is the accelerated development of the bulbs until anthesis or marketing of the plants. These phases have been discussed relative to floral and root development and the basic environmental requirements of the bulb species.