农杆菌介导三价融合基因Rirol转化八棱海棠的研究

研究了以口蹄疫病毒FMDV 2A序列融合多基因在八棱海棠遗传转化上的应用,将DREB,IRT1和rolC融合三价基因(Rirol)通过农杆菌介导法转化八棱海棠,获得了一批nptⅡ抗性植株,经报告基因检测、PCR和Southern杂交分析证明融合基因成功导入到八棱海棠中。对转基因八棱海棠相关性状分析表明,转基因八棱海棠比对照具有较强的耐盐性,在表型上,表现节间缩短,分枝性强,侧根发达,证明外源融合基因在转基因植株中得到表达。试验初步证明了利用FMDV 2A序列进行多基因转化八棱海棠的可行性,为苹果等果树多基因转化提供了新的思路和途径。     

超声波介导Mz_2NHX_1基因遗传转化提高珠美海棠耐盐性

【目的】提高珠美海棠的耐盐性,建立更高效的珠美海棠遗传转化体系。【方法】在转化液中利用超声波直接转化无菌条件下横切主脉的珠美海棠试管苗幼叶,通过愈伤组织培养诱导植株再生、抗性筛选,GUS染色鉴定,RT-PCR分析转基因植株的耐盐性。【结果】较利于珠美海棠愈伤组织诱导分化及抗性筛选的培养基为MS+6-BA 2.0 mg·L~(-1)+NAA 0.1 mg·L~(-1)+Kana 50 mg·L~(-1)。超声波处理的最优工作条件为:处理6 s,间歇10 s,工作重复20次,功率80 W,转化效率为31.3%。GUS染色与RT-PCR鉴定结果一致率为100%,RT-PCR结果分析显示,Mz2NHX1基因在转化苗中的表达量约为对照的3倍。荧光定量PCR结果显示转化苗Mz2NHX1基因的表达量为对照组的6.21倍,盐处理试验结果显示转化苗耐盐性得到了显著地提高。【结论】建立了超声波介导的珠美海棠高效遗传转化体系,获得珠美海棠耐盐新材料,为该耐盐基因的研究和盐碱地的改良奠定了基础。     

盐胁迫对不同耐盐性八棱海棠株系生理特性的影响

[目的]探究耐盐性不同的八棱海棠株系叶片生理指标对盐胁迫的响应模式,以期揭示耐盐性较强的八棱海棠株系的耐盐机制.[方法]采用盆栽过量灌溉盐水的方式,以耐盐性较强的八棱海棠(NY)组培株系为试材,普通八棱海棠(PT)组培株系为对照,以不同质量浓度NaCl(0、2、4、6、8、10 g·L-1)处理40 d,测定各处理植株...     

超声波辅助农杆菌介导八棱海棠转rolC基因

应用超声波辅助农杆菌介导法,对八棱海棠进行rolC基因转化,以期提高转化效率并获得转基因植株。利用gus基因瞬间表达的方法研究了超声波处理时间、处理时期和农杆菌悬浮液中乙酰丁香酮(As)浓度对rolC基因转化率的影响。结果表明,叶盘在D600nm为0．6且含有75 mg／L As的农杆菌悬浮液中侵染2 min后,用功率为100 W的超声波处理30 s,再浸泡2．5 min,然后放到再生培养基上共培养3 d,能获得最佳的gus基因瞬间表达率。最佳处理条件下转化683枚八棱海棠叶片,共得到138个抗性愈伤组织和15株抗性苗,转化率为2．2％。GUS染色、PCR及Southern blotting检测结果显示,有12个八棱海棠株系的基因组中整合了完整的外源rolC基因。     

五种苹果砧木的生长及生理特性对盐胁迫的响应

以八棱海棠(Malus robusta)、平顶海棠(Malus prunifolia)、红果海棠(Malus sieboldii)、珠美海棠(Malus zumi)、西府海棠(Malus micromalus)一年生实生苗为试材,对其进行盐胁迫处理,比较其耐盐性差异。结果表明:随盐浓度增加,参试植物的新梢生长量、叶面积和叶片厚度、根系活力降低,盐害率和盐害指数增加,其中八棱海棠的变化幅度最小,红果海棠相对较大;叶绿素(Chl)和类胡萝卜素(Car)含量变化各异,但八棱海棠和平顶海棠的变化较小;参试植株的可溶性糖及丙二醛(MDA)含量均随盐浓度增加呈上升趋势;超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均受不同程度影响,八棱海棠的SOD活性变化较小;八棱海棠的隶属函数平均值最大,红果海棠的最小。结合其形态表现综合分析认为,5种苹果砧木的耐盐性强弱依次为:八棱海棠、西府海棠、平顶海棠、珠美海棠、红果海棠。     

苹果砧木耐盐性田间鉴定

1997～ 1998年在山东省东营市盐碱地农场分别种植 14种 1年生苹果砧木苗和 6种砧木的1年生嫁接苗 ,发现砧木珠美海棠 ( Malus　 zumi)、小金海棠 ( M.xiaojinensis)耐盐性较强 ;M2 6、MM10 6、M9、八棱海棠 ( M.micromalus)耐盐性中等 ;丽江山定子 ( M.rockii)和 A2不耐盐。不同砧穗组合的耐盐能力主要取决于砧木 ,同一种砧木嫁接不同品种的耐盐性相当。轻度盐碱地育苗、覆膜铺草等栽培措施能提高苹果苗对盐胁迫适应性能力     

草木樨PUTNHX基因的转化及转基因苗的耐盐性

以草木樨破坏生长点的茎尖作为外植体,在含有不同浓度激素的分化培养基上诱导新生芽.获得最佳分化培养基配方MS+4 mg/L 6-BA+0.2 mg/L IAA.利用农杆菌侵染的方法进行PUTNHX基因的转化,获得大量转基因苗.在含盐量分别为0.4%、0.8%、1.2%的培养基中测定转化苗的耐盐性.结果表明:在0.8%浓度下,转化苗生长时间明显高于野生苗.野生苗第3周已全部死亡,而转化苗仅有个别叶片变黄,说明转基因苗可以耐0.8%的盐.PUTNHX基因的转化确实提高了草木樨的耐盐性.     

八棱海棠高效再生体系的建立及潮霉素的抗性试验

以八棱海棠子叶为外植体,比较了不同浓度植物激素、不同激素组合、不同暗培养时间及不同浓度潮霉素对八棱海棠再生体系的影响,建立了八棱海棠高效再生体系,筛选并分析了潮霉素在八棱海棠遗传转化中的适宜浓度。结果表明:诱导子叶愈伤的最佳培养基为Sc+2.0 mg·L~(-1) TDZ+0.2 mg·L~(-1) NAA+0.5 mg·L~(-1) GA_3,芽再生的最佳培养基为Sc+2.0 mg·L~(-1) TDZ+0.2 mg·L~(-1) NAA,暗培养最佳时间为10 d,生根最佳培养基为1/2MS+0.25 mg·L~(-1) IBA。八棱海棠在整个再生过程中均对潮霉素敏感,在潮霉素浓度为2.0 mg·L~(-1)的愈伤诱导培养基上,外植体的愈伤诱导率仅为23.33%,分化率仅为9.88%;当潮霉素浓度为4.0 mg·L~(-1)时,愈伤诱导率下降为13.00%,而且愈伤不分化,随着培养时间延长,愈伤组织变得更加致密,之后变褐死亡。在诱导生根培养基上添加3.0 mg·L~(-1)潮霉素,生根率仅为3.52%,大部分外植体再生芽底部只会诱导出愈伤组织,没有根原基形成。八棱海棠子叶愈伤诱导及分化的潮霉素筛选临界浓度为2.0～4.0 mg·L~(-1),生根时潮霉素筛选浓度为2.0～3.0 mg·L~(-1)。     

一种快速筛选转抗霜霉病基因甜瓜种子的方法研究

以用花粉管通道技术转入抗霜霉病基因At2的甜瓜T1代转基因种子为试材,采用组培技术得到膨大的外植体,利用PCR技术对组培得到的外植体进行检测,以期尽快筛选出阳性植株。结果表明:在甜瓜外植体培养过程中,筛选到的种子最佳灭菌条件为72℃、干灭4h,抑菌效果最好的培养基组分是MS无机盐+B5维生素+2mg/L 6-BA+7g/L琼脂+500mg/L头孢噻肟钠+1.1g/L甲基托布津;对1 100个转抗霜霉病基因At2的T1代种子外植体进行PCR检测,阳性率为4.5‰。     

三种苹果属植物幼苗拒Na2+机理的研究

 以苹果属耐盐性不同的山定子、小金海棠和珠眉海棠的幼苗为试材，通过拒Na²⁺ 能力的测定确定了盐敏感种的整体拒Na²⁺ 能力为65%左右，中等耐盐种的整体拒Na²⁺ 能力为80%左右，耐盐种的整体拒Na 能力为90%左右，根部和茎基部的拒Na 能力是苹果属植物幼苗整体拒Na²⁺ 能力的主要部分，耐盐种根部和茎基部的拒Na²⁺ 能力明显大于盐敏感种。耐盐种侧根、茎基木质部和老叶的Na²⁺累积效应明显，是主要的聚Na²⁺ 部位。耐盐种通过地上部Na²⁺ 的重新分配和再转运，使幼叶和成熟叶Na²⁺ 含量更低。     

Effects of radiation from 188Re on smooth muscle cell proliferation

AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β^-particles emission from ¹⁸⁸Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by ¹⁸⁸Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [³H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of ¹⁸⁸Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from ¹⁸⁸ Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G₀/G₁ arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.     

Effect of L-Arg on plasma content of endothelin and expression of c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy

AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration.     

Ergebnisse der Leistungsprüfungen der Süßkirschensorte ‚Stella‘ auf Gisela- und Weiroot-Unterlagen in Bulgarien

Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden.     

Compositional and Functional Properties of Saskatoon Berry and Blueberry

Abstract

Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared.     

多效唑对猕猴桃离体试管苗生长及内源激素的影响

多效唑（ＰＰ３３３）处理猕猴桃试管苗，降低了其生长强度；植株体内的ＧＡ３、ＩＡＡ和ＺＴ含量下降，ＡＢＡ的含量上升，乙烯释放率增加；并且能降低外源的ＧＡ３和ＩＡＡ促进生长的作用，而外源的ＧＡ３和ＩＡＡ又能不同程度地逆转多效唑的抑制作用，使植株恢复生长。     

Proteomic analysis between pathological scars and normal skin

AIM: To investigate and screen the sensitive proteins in the formation mechanism of pathological scars by comparing the results of differential proteomic analysis between pathological scars and normal skin.METHODS: Two-dimensional gel electrophoresis was used to detect the protein expression profiles in 8 keloid patients, 8 hypertrophic scar patients and 3 matched normal skin patients.The proteins that showed differential expression of over 4-fold change were cut and analyzed by MALDI-TOF/TOF mass spectrometry.RESULTS: A two-dimensional protein profiling comparison between pathological scars and normal skin was successfully established.On average, 2 978 spots in keloid, 2 975 spots in hypertrophic scar and 3 053 spots in normal skin were identified using gel analysis software.Compared with normal skin, there were totally 36 differentially-expressed proteins in keloid and hypertrophic scar identified from the spots of over 4-fold change, including 16 proteins in both keloid and hypertrophic scar (8 up-regulated and 8 down-regulated), 11 only in keloid (9 up-regulated and 2 down-regulated) and 9 only in hypertrophic scar (4 up-regulated and 5 down-regulated).CONCLUSION: Proteomic analysis can identify the proteins with variance of pathological scars versus normal skin, thus providing probable new clues to reveal the formation mechanism of pathological scars.     

Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.     

Metallothionein inhibits homocysteine-induced proliferation of rat vascular smooth muscle cells

AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [³-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10^-6-10^-4 mmol/L) stimulated [³-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [³-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10^-6-10^-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl₂ for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.     

Coupling past management practice and historic landscape change on John F. Kennedy Space Center,Florida

Historic landcover dynamics in a scrubby flatwoods (Tel-4) and scrub landscape (Happy Creek) on John F. Kennedy Space Center were measured using aerial images from 1943, 1951, 1958, 1969, 1979, and 1989. Landcover categories were mapped, digitized, geometrically registered, and overlaid in ARC/INFO. Both study sites have been influenced by various land use histories, including periods of range management, fire suppression, and fire management. Several analyses were performed to help understand the effects of past land management on the amount and spatial distribution of landcover within the study sites. A chi-squared analysis showed a significant difference between the frequency of landcover occurrence and management period. Markov chain models were used to project observed changes over a 100-year period; these showed current management practices being effective at Tel-4 (restoring historic landscape structure) and much less effective at Happy Creek. Documenting impacts of past management regimes on landcover has provided important insight into current landscape composition and will provide the basis for improving land management on Kennedy Space Center and elsewhere.     

XBP-01 accelerated apoptosis induced by oxidized low density lipoprotein in vascular smooth muscle cells

AIM: Previous studies performed with XBP-01 in vitro indicated that XBP-01 could inhibit vascular smooth muscle cells from being transformed into foam cell and could eliminate the atherosclerotic plaque in C57BL/6J mouse. This experiment is to investigate its mechanism of eliminating plaques in vitro. METHODS: The cultured porcine artery smooth muscle cells incubated with XBP-01 of 0.1 mg/L for 24 h after preincubated with oxidized low density lipoprotein of 15 mg/L for 72 h in vitro. The samples were analyzed by fluorescence microscope, confocal microscope system and flow cytometry. RESULTS: Apoptosis was triggered by being incubated with oxidized low density lipoprotein and this process was accelerated additionally by being incubated with XBP-01. CONCLUSION: XBP-01 can be effective in eliminating atherosclerotic plaque by accelerating the process in which oxidized low density lipoprotein induced smooth muscle cell apoptosis.