木薯MeSAUR1互作蛋白的筛选及鉴定

木薯MeAGPS1a基因编码的小亚基是淀粉合成关键酶AGPase的催化中心,前期研究发现MeSAUR1转录因子可结合MeAGPS1a基因启动子并调控该基因表达。采用酵母双杂交技术筛选MeSAUR1转录因子的互作蛋白,可进一步解析MeAGPS1a基因表达的分子调控网络。本研究构建了MeSAUR1的酵母双杂交诱饵载体pGBKT7-MeSAUR1,采用酵母双杂交技术从木薯cDNA文库中筛选MeSAUR1的候选互作蛋白,结果共筛选出31个阳性克隆。通过酵母双杂交点对点实验验证了候选互作蛋白MePP2C与MeSAUR1的互作关系,本研究结果有助于揭示Me SAUR1蛋白调控木薯淀粉合成的互作网络。     

红掌AaMYB1的表达特征及其在烟草中的过表达

R2R3-MYB转录因子在观赏植物花青素着色中具有关键性的调控作用。为了进一步明确红掌AaMYB1调控花青素合成的功能,本研究采用序列分析、表达特征分析以及在烟草中异源过表达等方法,对AaMYB1进行了功能分析。结果表明,AaMYB1与单子叶植物中调控花青素合成的R2R3-MYB同源性较高。在品种‘Tropical’中,AaMYB1主要在红掌佛焰苞中表达,各组织中的表达量与花青素苷积累密切相关,但在不同品种不同颜色佛焰苞中的表达与花青素苷积累无相关性。单独过表达AaMYB1的T1代烟草花冠檐部因大量积累花青素苷而使颜色变深,且烟草中花青素苷合成途径上的关键酶基因NtDFR和NtANS以及烟草内源调控花青素苷合成的R2R3-MYB基因NtAn2的表达量得到大幅度上调。以上结果进一步明确了AaMYB1具有调控花青素苷合成的功能。     

‘月月粉’和野蔷薇花青素合成酶基因的鉴定与表达分析

植物花青素是一种天然的黄酮类水溶性植物色素,对于植物的花和果实的花色决定具有重要作用,花青素在植物体内的生物合成途径研究相对较清楚。但在蔷薇属植物中花青素合成酶基因的功能分析相对较少,通过序列比对和进化树分析鉴定‘月月粉’(Rosa chinensis‘Old Blush’,用OB表示)和野蔷薇(Rosa multiflora)中花青素合成酶基因各14个。通过序列比较发现野蔷薇和OB中对应的同源基因序列相似性很高,表明这些基因的保守性很强,但表达谱分析显示这些基因在两个物种中的差异较大,暗示其调控序列可能变异程度较大。通过检测一个野蔷薇自然突变单株中红花和白花中的花青素合成酶基因的表达,推测有部分花青素合成酶基因可能在花青素合成反馈调节中具有重要作用。通过分析启动子序列中顺式作用元件推测OB中花青素合成酶可能受蔗糖和不同激素的调控。本研究可为揭示蔷薇属植物花青素合成调控的分子机制提供依据。     

‘红颜’草莓果实黄烷酮3-羟化酶基因克隆及表达分析

为探究八倍体‘红颜’草莓(Fragaria×ananassa Duch.)果实着色过程中的关键基因,本研究通过对前期果实表皮部位RNA-Seq数据的研究,筛选花色素苷合成途径的差异表达基因黄烷酮3-羟化酶基因(Flavanone 3-hydroxylase)。根据‘红颜’草莓F3H基因序列设计一对引物,利用RT-PCR技术从中克隆F3H核苷酸序列,命名为FaF3H。将基因上传GenBank数据库获得八倍体‘红颜’草莓F3H基因登录号(MZ190021),并进行生物信息学分析。利用实时荧光定量方法分析草莓果实着色过程中该基因表达水平。结果显示,‘红颜’草莓FaF3H基因长度为1 095 bp,编码364个氨基酸。保守结构域分析显示,具有一个PLN02515超家族结构域。RT-qPCR结果表明FaF3H基因在草莓果实大绿期到白果期表达量下降,全红期表达量最高。FaF3H基因在草莓果实花色素苷合成过程中发挥重要作用。     

苹果MdPIF4基因克隆、表达与功能验证

温度是影响植物生长发育重要的环境因子之一,光敏色素互作因子PIF4是植物响应温和高温的核心基因,明确MdPIF4在苹果温度响应中的作用具有重要意义。本研究克隆了MdPIF4的编码区和启动子序列,分析了MdPIF4的表达情况,对MdPIF4转录激活活性进行了分析和互作蛋白筛选,并在野生型拟南芥中异源过表达了MdPIF4。结果显示:MdPIF4基因编码区包含1 656 bp的核苷酸,编码551个氨基酸,预测其蛋白质分子质量为60 670.87 Da,等电点为6.63。MdPIF4在苹果叶片中的表达量最高,其次为根、茎、花,在果实中的表达量最低。MdPIF4启动子中含有光响应元件、ABA(脱落酸)响应元件ABRE、SA (水杨酸)响应元件TCA-element、厌氧调节响应元件ARE、干旱响应元件MBS等。MdPIF4受温和高温诱导表达。不同物种间bHLH结构域的蛋白序列保守性较强,MdPIF4序列全长在酵母中具有转录自激活活性,bHLH结构域在酵母中没有自激活活性,其余片段均具有自激活活性。以bHLH结构域为诱饵蛋白,酵母双杂交筛库获得3个互作蛋白,分别为Md WRKY70、MdHAT5和MdRab11D。在野生型拟南芥中过表达MdPIF4促进了拟南芥幼苗下胚轴的生长。上述研究表明MdPIF4参与苹果温和高温响应。     

枣ZjERF53转录因子的转录激活分析及互作蛋白筛选

乙烯应答因子广泛参与调控植物生长发育和非生物胁迫响应过程。本研究克隆了1个在枣(Ziziphus jujuba Mill.)果实成熟后期高表达的ZjERF53转录因子,对其进行了亚细胞定位、转录激活活性和互作蛋白筛选研究,以揭示其生物学功能。研究结果表明：ZjERF53基因编码区序列全长为1 155 bp,能在细胞核和细胞膜中表达;ZjERF53具有自激活活性,其转录激活功能区域位于蛋白序列C端;以无自激活活性pGBKT7-ZjERF53-N为诱饵,通过酵母双杂交技术筛选枣果实cDNA文库,共获得4个互作蛋白。本研究揭示了ZjERF53可能参与枣树抗病和抗旱等非生物胁迫响应,为解析ERF参与调控枣树生物学过程的分子机制提供了工作基础。     

苦荞FtHY5基因克隆、生物信息学及参与花青素合成分析

HY5 (Elongated hypocotyl 5)转录因子是参与植物光形态建成的重要成员,并调控花青素的生物合成。本研究以苦荞‘晋荞2号’作为材料,利用PCR克隆其HY5基因的完整的CDS序列,并对FtHY5的理化性质、保守功能域、信号肽、亚细胞定位、亲/疏水性、蛋白质结构、系统进化等进行了生物信息学分析,此外还分析了其对苦荞芽菜花青素合成的影响。结果表明,FtHY5基因完整的CDS长度为507 bp,编码的蛋白分子量为18.434 kD,等电点为9.47,空间构象呈拉链形状,属于亲水性蛋白,含有信号肽,亚细胞定位在细胞核里。苦荞FtHY5蛋白的氨基酸序列与藜麦和甜菜的亲缘关系最近。FtHY5基因与花青素生物合成途径上的结构基因在光照条件下培养的芽菜中的表达水平均高于黑暗条件下的,说明Ft HY5可能调控这些花青素合成基因的表达水平,从而参与光诱导花青素合成的过程。综上所述,本研究结果将为进一步研究FtHY5调控苦荞芽菜花青素的生物合成的分子机理提供基础,同时也为利用该基因来改善苦荞芽菜的品质提供理论支撑。     

银杏FLS基因启动子克隆及序列分析

黄酮醇合成酶基因（FLS）是黄酮类化合物合成通路中的重要基因,直接决定着银杏黄酮类化合物的积累量。为了探索FLS基因在转录水平上的表达调控机制,本文在克隆银杏FLS基因启动子序列的基础上,利用生物信息学方法,分析了该基因的启动子结构。结果表明,FLS基因启动子中包含了3个MYB转录因子结合位点和9个光信号应答位点,为确立银杏FLS基因与CHI基因间的共调控关系及揭示FLS基因应答光信号的转录调控机制提供了分子基础。     

影响葡萄果皮颜色的MYB类转录因子的研究进展

在葡萄品种中,果皮颜色从白色到黑色存在广泛的差异,花青素苷的合成和积累是形成果皮颜色的直接原因,MYB类转录因子是调控花青素苷合成的一类重要的转录因子。为了进一步探究不同种果皮颜色同MYB转录因子之间的关系,本研究列出了花青素的结构和种类以及花青素苷生物合成途径,归纳了葡萄色泽与花青素苷的关系,列举了调控植物花青素苷合成的相关基因的功能,并着重分析了葡萄MYB类转录因子在花青素苷合成过程中的功能,以期为今后改良葡萄果皮颜色提供指导。     

Ethanol vapor and saprophytic yeast treatments reduce decay and maintain quality of intact and fresh-cut sweet cherries

The objective of this study was to evaluate the use of an ethanol vapor release pad and a saprophytic yeast Cryptococcus infirmo-miniatum (CIM) to reduce decay and maintain postharvest quality of intact or fresh-cut sweet cherries (Prunus avium) cv. Lapins and Bing. Intact or fresh-cut fruit were packed in perforated clamshells (capacity 454 g) and stored at 1, 10 or 20 °C for up to 21, 14 and 8 d, respectively. For ethanol treatment, a pad made with silica gel powder containing 10 g ethanol and covered with perforated film, which allows ethanol vapor to diffuse gradually, was attached to the upper lid of the clamshells. Ethanol treatment caused accumulation of ethanol in the packaging headspace, about 10 μL L⁻¹ with little change within 14 d at 1 °C, 23 μL L⁻¹ at d 1 and decreased to 15 μL L⁻¹ at d 10 at 10 °C, and 26 μL L⁻¹ at d 1 and decreased to 13 μL L⁻¹ at d 3 at 20 °C. Ethanol content in fruit was less than 9 mg kg⁻¹ in all the control fruit, and increased to 16, 34 and 43 mg kg⁻¹ in ethanol-treated fruit at 1, 10 and 20 °C, respectively. Nonetheless, a sensory taste panel did not perceive any flavor difference from the ethanol treatment. The ethanol treatment retarded softening, darkening, and acid decrease in fruit as well as discoloration of the stems, and extended shelf-life of intact cherries. Ethanol reduced brown rot (Monilinia fructicola) in fresh-cut cherries stored at 20 °C, but not at 1 and 10 °C. A pre-packaging dip in CIM completely controlled brown rot in inoculated fresh-cut cherries stored at 1 °C, and in naturally infected cherries at 20 °C.     

膜下滴灌条件下高产甜菜灌溉的生理指标

甜菜是我国重要的糖料作物,其生物产量高,需水量大,合理灌溉是节约用水、提高产量的有效措施之一。本试验连续两年研究了内蒙古半干旱地区膜下滴灌条件下,不同灌水量甜菜块根产量与叶面积指数、净光合速率、蒸腾速率、叶水势、土壤含水量和耗水量之间的关系,以及不同灌水量对甜菜产量和水分利用效率的影响。结果表明,高产甜菜的叶面积指数在叶丛快速生长期大于7.37,在块根糖分增长期和糖分积累期分别为6.08~6.51和4.19~5.57,在叶丛快速生长期、块根糖分增长期和糖分积累期叶水势分别为–0.09~–0.22、–0.18~–0.39和–0.26~–0.48 MPa,净光合速率分别为21.28~28.23、21.90~28.75和22.06~26.58μmol m–2 s–1,蒸腾速率在叶丛快速生长期和块根糖分增长期分别为9.36~10.21 mmol m–2 s–1和6.37~7.73 mmol m–2 s–1,在糖分积累期大于4.69 mmol m–2 s–1,耗水量分别为140.15~312.78、44.93~200.45和56.32~113.06 mm。甜菜产量、产糖量、水分利用效率均高的合理灌溉量,在丰雨年份(生育期降雨量500 mm)为1350 m3 hm–2,在少雨年份(生育期降雨量300 mm)为1800 m3 hm–2,为甜菜节水灌溉提供了理论依据和生理指标。     

The effect of temperature and other factors on chlorophyll a fluorescence and the lower oxygen limit in apples (Malus domestica)

The effects of temperature, scan interval and rate of oxygen (O₂) decline on pulse frequency modulation (PFM)-based minimum fluorescence (F_α) and the F_α-based lower oxygen limit (LOL) were investigated using ‘Honeycrisp’ apples (Malus × domestica Borkh). The effects of temperature and hypoxic stress on pulse amplitude modulation (PAM) fluorescence parameters were also investigated. The PFM scan interval had no effect on the F_α baseline, but increases in the scan interval decreased the low-O₂-induced fluorescence spike intensity (ΔF_α). Temperature negatively correlated with the F_α baseline while ΔF_α °C⁻¹ was greater at lower than at higher temperatures. When using a PAM fluorometer, the minimum fluorescence (F_o), and to a lesser extent the maximum fluorescence (F_m), were similarly affected by temperature as was F_α. Temperature altered the LOL in fruit. Apples stored at 20, 10, 3.5 and 0 °C spiked at 0.72, 0.33, 0.22 and 0.08 kPa O₂, respectively, under a rapid O₂ decline (i.e., 20.9 to <1 kPa O₂ in ≈5–6 h). Although the low-O₂ F_α spike apex values did not change with temperature, the spike intensity increased with temperature due to a reduced fluorescence baseline. A slower O₂ decline rate produced slightly higher LOL and lower spike intensity values. In conclusion, temperature and rate of O₂ decline affected the low-O₂-induced PFM fluorescence spike intensity as well as the LOL, while the PFM scan interval affected the spike intensity.     

Effect of two wheat genotypes and Swedish environment on falling number, amylase activities, and protein concentration and composition

Variation in falling number, amylase activities, protein concentration and composition were investigated in two wheat cultivars grown in Sweden over two seasons, in four locations, with four N fertilizer rates, with and without fungicide treatment. The results showed that;

Cultivar differences in gaseous 1-methylcyclopropene accumulation in whole and fresh-cut apple fruit

A number of studies have shown that responses of apple fruit to 1-methylcyclopropene (1-MCP) vary considerably among cultivars. This study was designed to determine if cultivars show differences in accumulation of gaseous 1-MCP. Apple fruit were placed in 1.76 L jars that were sealed and injected with 20 μL L⁻¹ 1-MCP. After 12 h, samples of intercellular atmosphere were removed and analyzed for 1-MCP concentration. Accumulation of internal gaseous 1-MCP varied markedly among cultivars, ranging from 0.14 ± 0.06, 0.22 ± 0.03, and 0.77 ± 0.30 in ‘Redcort’, ‘McIntosh’, and ‘Empire’, respectively, to 2.10 ± 0.28, 3.33 ± 0.13, and 6.93 ± 0.35 μL L⁻¹ in ‘Gala’, ‘Cameo’, and ‘Honeycrisp’, respectively. Accumulation of gaseous 1-MCP was reduced an average of 51% in fruit treated with Sta-Fresh 8711 fruit wax. The role of the epidermis in modulating 1-MCP ingress was determined by measuring gaseous 1-MCP accumulation in fresh-cut tissue. Fresh-cut cortical tissue rapidly depleted headspace 1-MCP (>95%) over a 1-h exposure yet accumulated negligible quantities of internal gaseous 1-MCP. By contrast, cortical tissue treated with ascorbic acid or hypotaurine, or aged for several hours prior to exposure to 1-MCP, showed reduced consumption of headspace 1-MCP and high accumulation of internal gaseous 1-MCP. Levels of internal 1-MCP in cortical tissue from the cultivars generally paralleled those for intact fruit, ranging from 0.23 ± 0.07, 0.37 ± 0.18 and 1.09 ± 0.14 μL L⁻¹ in ‘Empire’, ‘McIntosh’ and ‘Redcort’, respectively, to 2.40 ± 0.71, 4.55 ± 0.15, and 6.24 ± 0.85 in Gala’, ‘Cameo’, and ‘Honeycrisp’, respectively. Although commercial fruit wax influences gaseous 1-MCP accumulation, the comparable accumulation patterns in unwaxed whole and fresh-cut apple fruit suggest that epidermal tissue/native waxes alone do not account for cultivar differences.     

Inheritance and allelic relationship among gene(s) for blast resistance in pearl millet [Pennisetum glaucum (L.) R. Br.]

Six blast‐resistant pearl millet genotypes, ICMB 93333, ICMB 97222, ICMR 06444, ICMR 06222, ICMR 11003 and IP 21187‐P1, were crossed with two susceptible genotypes, ICMB 95444 and ICMB 89111 to generate F₁s, F₂s and backcrosses, BC₁P₁ (susceptible parent × F₁) and BC₁P₂ (resistant parent × F₁) for inheritance study. The resistant genotypes were crossed among themselves in half diallel to generate F₁s and F₂s for test of allelism. The F₁, F₂ and backcross generations, and their parents were screened in a glasshouse against Magnaporthe grisea isolates Pg 45 and Pg 53. The reaction of the F₁s, segregation pattern of F₂s and BC₁P₁ derived from crosses involving two susceptible parents and six resistant parents revealed the presence of single dominant gene governing resistance in the resistant genotypes. No segregation for blast reaction was observed in the F₂s derived from the crosses of resistant × resistant parents. The resistance reaction of these F₂s indicated that single dominant gene conferring resistance in the six genotypes is allelic, that is same gene imparts blast resistance in these genotypes to M. grisea isolates.     

干旱荒漠区物理结皮的土壤水文效应

在干旱荒漠区,物理结皮是广泛发育的一类结皮,其存在对维持干旱荒漠区生态平衡具有重要作用。笔者总结了物理结皮种类、形成及影响因素,综述了物理结皮对土壤降雨入渗、蒸发、地表径流以及土壤发育和微生物生长等方面的国内外研究动态,探讨了物理结皮和生物结皮的相互联系,在此基础上,提出了物理结皮进一步研究的展望,认为：在河西走廊东段的民勤沙区,粘土沙障加梭梭的固沙造林模式,促使粘土沙障在雨滴的冲刷下物理结皮广泛发育,发育的结皮降低了降水入渗,增加了地表径流,使深层土壤旱化,造成人工梭梭固沙林衰退和较深层土壤水分的减少,导致沙区生态水文过程和植被格局变化,研究对退化荒漠植被恢复与干旱沙区土壤水分循环具有意义。     

Ecuador, 1991 potato germplasm collecting expedition: taxonomy and new germplasm resources

Summary We conducted a joint Ecuador/Colombia/United States wild potato (Solanum sect. Petota) germplasm collecting expedition in Ecuador from April 13–July 1, 1991. The goals of the expedition were to collect germplasm and study the species boundaries of all of the 25 Ecuadorian taxa accepted by current taxonomists. We made 126 collections of 24 of these 25 taxa, 113 as germplasm samples, 13 only as herbarium collections. We synonymize six of these 25 names (S. baezense Ochoa, S. cyanophyllum Correll, S. pichinchense Bitter & Sodiro, S. serratoris Ochoa, S. suffrutescens Correll as synonyms of S. andreanum Baker; S. correllii Ochoa as a synonym of S. regularifolium Correll). Four other names (S. chomatophilum f. angustifoliolum Correll, S. moscopanum Hawkes, S. solisii Hawkes, S. tundalomense Ochoa) could not be consistently distinguished from S. colombianum Dunal in the field. We are currently investigating them to determine their species status.     

Performance of various grapefruit (<Emphasis Type="Italic">Citrus paradisi</Emphasis> Macf.) and pummelo (<Emphasis Type="Italic">C. maxima</Emphasis> Merr.) cultivars under the dry tropic conditions of Mexico

Grapefruit growers in the tropics require information about existing and new citrus cultivars with high productivity potential. The objective of this study was to determine the growth, yield, and fruit quality performance of seven pigmented and four white grapefruit cultivars under the dry tropic conditions of Colima, Mexico. The trees were budded on sour orange (Citrus aurantium L.) rootstock and planted at a distance of 8 × 4 m. ‘Oroblanco’ and ‘Marsh Gardner’ white-fleshed grapefruit cultivars and ‘Chandler’, a pink-fleshed pummelo, were the largest trees with the greatest height (5.0–5.6 m), canopy diameter (6.2–6.3 m), trunk diameter (21.9–23.3 cm), and canopy volume (109–123 m³). Lower height (4.3–4.8 m) and canopy volume (73–96 m³), but with similar canopy diameter to the previously mentioned cultivars, were recorded for the remaining pigmented cultivars. ‘Chandler’ pummelo and four pigmented grapefruit cultivars (‘Shambar’, ‘Río Red’, ‘Ray Ruby’, and ‘Redblush #3’) had yearly productions of 34.8, 34.9, 34.1, 32.7, and 30.6 ton ha⁻¹, respectively. The most productive white grapefruit cultivar was ‘Marsh Gardner’ (30.5 ton ha⁻¹). Grapefruit cultivars having the largest fruit size showed a higher inverse relationship between fruit weight and yield than those with small fruit. Most genotypes had higher values of fruit weight, juice content, and maturity index than those required by the local market. The most promising grapefruit cultivars based on their acceptable growth, yield superior to 30 ton ha⁻¹, and acceptable fruit color were ‘Río Red’, ‘Shambar’, ‘Ray Ruby’, and ‘Redblush #3’.     

Structural and physiological changes associated with the skin spot disorder in apple

Skin spot is an important physiological disorder of ‘Elstar’ apples (Malus × domestica Borkh.) that occurs after fruit have been removed from controlled atmosphere storage. Skin spots are irregular patches of small, round, brown blemishes. Cross-sections reveal a browning of protoplasts (coagulated) and of cell walls that extends into the hypodermis. Skin spots are always associated with linear, gaping and non-gaping microcracks in the cuticle. Staining of apple skin with calcofluor white usually results in white fluorescence of cell walls but, within a skin spot, the white fluorescence is weak or absent. Cell walls within, and in the immediate vicinity of skin spots stain with phloroglucin/HCl indicating the presence of lignin. The area of skin affected by skin spots was positively and linearly correlated with the area of the non-blush fruit surface infiltrated by acridine orange. In general, skin spots were limited to the non-blush fruit surface and occurred more frequently near the stem-end than the calyx region of the fruit. Skin spot areas were correlated with a 2.5-fold increase in water vapour permeability compared with unaffected areas (23.8 ± 4.0 m s⁻¹ with skin spots, 9.6 ± 2.1 × 10⁻⁵ m s⁻¹ without skin spots). Strips of the fruit skin from regions with skin spots had an increased maximum force and modulus of elasticity. Dipping fruit in ascorbic acid (0.1 or 0.3 mM for 10 min) before storage decreased the area affected by skin spots. There was no effect of dipping in ethanol/water (70%, v/v, 15 min) or in solutions of captan (1.5 g L⁻¹, 10 min) or trifloxystrobin (0.1 g L⁻¹, 10 min). In contrast, prestorage treatment with 1-methylcyclopropene (630 nL L⁻¹ for 24 h) or poststorage incubation in H₂O₂ (10% for 2, 6, 10 and 16 h) increased skin spots. Our data are consistent with a typical cell response to an oxidative burst that seems to be focussed on particular regions of the ‘Elstar’ fruit surface by concentrations of cuticular microcracks, and that is possibly caused by reoxygenation injury upon removal from CA storage.