莴苣高效瞬时表达体系的建立

 以GUS基因作为报告基因, 利用农杆菌真空渗透法, 采取正交试验设计对莴苣(Lactuca sativa L. sp) 的瞬时表达条件进行研究, 建立了高表达水平的瞬时转化体系(乙酰丁香酮200 μmol /L、菌液密度OD₆₀₀值0.8、真空处理30 min、共培养6 d) 。应用结果表明: 优势组合的表达水平比对照的表达水平提高16.3倍。     

基因瞬时表达技术在园艺植物上的应用研究进展

基因瞬时表达技术是近年来发展形成的一种快速高效的基因功能分析方法,应用广泛.本文综述了基因瞬时表达技术的原理、影响因素及其在园艺植物上的应用,指出该技术在当前应用中存在的不足并提出建议及展望.     

基因瞬时表达技术在园艺植物上的应用研究进展

 基因瞬时表达技术是近年来发展形成的一种快速高效的基因功能分析方法,应用广泛。本文综述了基因瞬时表达技术的原理、影响因素及其在园艺植物上的应用,指出该技术在当前应用中存在的不足并提出建议及展望。     

FaASR基因超表达促进草莓果实着色

以‘甜查理’草莓为试材,在克隆草莓ASR同源基因的基础上,对草莓中FaASR基因进行生物信息学、时空表达分析,通过调控草莓果实中ASR基因表达水平,分析果实的着色、蔗糖、ABA、色素和果实硬度等生理指标的变化,以及与色素代谢相关基因CHS、UFGT的表达水平,揭示ASR基因调控草莓果实成熟的机制。草莓ASR含有一个典型的与果实成熟和抗逆有关的ABA/WDS区域,ASR基因过量表达可以促进草莓果实提前上色,促进果实内源ABA积累、蔗糖含量增加和果实变软,并且促进与果实花色苷积累相关的关键基因CHS和UFGT的转录水平。该研究有助于深入揭示果实发育信息传递的分子机制,也为今后草莓的分子改良育种奠定重要的基础。     

根癌农杆菌介导外源基因转化番茄体系优化

为建立优化的番茄外源基因转化体系,提高番茄的外源基因转化率,以番茄叶盘预培养时间和农杆菌与番茄叶盘的共培养温度、共培养时间为试验要素,摸索相对适宜的预培养及共培养的时间和温度范围,其余操作按常规步骤进行。结果表明:农杆菌与番茄叶盘的预培养适宜时间为27℃下32～48h;农杆菌与番茄叶盘的共培养的最适宜条件是:22～24℃下暗培养48h。     

草莓果实的消费市场探讨

通过“草莓品尝问卷调查”,就草莓果实的商品性状与消费者的心理、习惯、行为及消费群体等,进行调查统计、作图分析与综合归类。结果显示:消费者喜欢的品种是达赛莱克特、土特拉和卡图诺,它们的色泽鲜艳浓红,果形长圆锥形,酸甜适度;但丰香在芳香和口感上仍有优势。消费者的群体结构主要在年龄56岁以下,家庭收入为中等水平(3万～5万元/年),尤其是女性;受文化教育程度越高,对草莓的偏爱越深。     

根癌农杆菌介导的韭菜基因转化体系的建立

 以GUS 基因为报告基因, 建立了农杆菌介导的韭菜基因转化体系。研究结果表明, 以预培养4～6 d 的根尖为转化受体, 用OD₆₀₀0. 6 ～0. 8 的LBA4404 浸染2～5 min , 共培养2 d , 然后在MS + NAA1 mg/L + BA 2 mg/L + Km 25 mg/L + Timentin 400 mg/L + As 100 mg/L 培养基上培养40 d , 后有愈伤组织产生,转入光下培养20 d 后有少量绿色不定芽产生。经X-Gluc 染色、PCR 分析和Southern blot 检验, GUS 基因已经整合到韭菜基因组染色体上。     

草莓果实营养成份的分析

草莓果实营养成份的分析万清林，赵书清（哈尔滨师范大学生物系）（黑龙江大学化学系）草莓是营养丰富的水果，在世界上的栽培面积较广，占小浆果栽培面积的首位，熟期在６—７月份，可以补充水果淡季市场、近年来黑龙江省的栽培面积也在不断扩大，其营养价值也为人们所逐...     

农杆菌介导TCS及RIP基因转化草莓的研究

采用农杆菌介导法,将TCS基因和RIP基因导入草莓中,探讨影响草莓遗传转化的主要因素,建立了高效的草莓遗传转化体系。并经Km抗性筛选,获得了若干转基因植株,对这些植株进行了TCS基因和RIP基因的PCR检测和PCR-Southern杂交检测,结果显示,TCS基因及RIP基因已转入草莓植株的基因组中。     

黄瓜疏松愈伤组织瞬时表达系统的建立

在NH4NO3浓度为2.0 g/L、6-BA浓度为1.0 mg/L、NAA浓度为0.1 mg/L的MS培养基中添加酵母提取物5 g/L有利于产生质地疏松的愈伤组织,适合作为外源基因瞬时表达的受体。     

Heat treatments and expansin gene expression in strawberry fruit

Heat treatments have been applied in fruit postharvest technology for insect disinfestations, decay control, ripening delay and modification of fruit responses to other stresses. Heat treatment affects several aspects of fruit ripening, such as ethylene production and cell wall degradation probably through changes in gene expression and protein synthesis. In this paper, strawberries (Fragaria × ananassa Duch., cv Camarosa) at 50–75% red stage of ripening were heat-treated at 45 °C during 3 h in an air oven and then stored at 20 °C for 0, 4, 18, 24 and 48 h. Fruit firmness was determined and the expression of a set of expansin genes (FaEXP1, FaEXP2, FaEXP4, FaEXP5 and FaEXP6) was analyzed. The firmness of treated fruit was higher than that of control fruit 24 h after treatment, though the differences disappeared after 48 h at 20 °C. The analysis by northern-blot indicated that heat treatments affected differently the expression of expansin genes. The expression of FaEXP1, FaEXP2 and FaEXP6 was lower in treated fruit during the following 24 h post-treatment. The lower expression of these expansin genes could contribute to delay softening after heat treatment.     

草莓果实发育过程中糖、pH值及ABA水平变化趋势

为了揭示草莓果实发育和成熟的生理机制,本研究以杜克拉草莓7个时期的果实为试材,分析果实发育过程中pH值、可溶性糖和ABA变化趋势.结果表明,果实发育前期可溶性糖积累以葡萄糖和果糖为主,蔗糖很低;果实进入始熟和成熟,蔗糖分别有一次快速积累的过程,全红时蔗糖含量是果糖和葡萄糖的1.25倍.以纯白期pH值最低点为界,pH值先...     

全基因组基因表达分析技术及其在果树上的应用

基因表达分析对于揭示基因功能、了解基因间相互关系、阐明植物生长发育过程中的生理生化调控机制起着至关重要的作用。综述了mRNA差异显示、cDNA-AFLP、基因芯片、EST数据分析、基因表达系列分析(SAGE)等全基因组基因表达分析技术的原理特点及在果树上的应用情况,并介绍了利用全基因组基因表达分析技术开展果树研究的思路。     

Fruit yield and quality of strawberry plants transformed with a fruit specific strawberry pectate lyase gene

Two transgenic strawberry lines (Pel 1 and Pel 3) containing the open reading frame of a fruit specific strawberry pectate lyase gene (FaplC) under the control of the CaMV35S promoter have been obtained to evaluate the role of this gene on fruit softening. Ripen fruits from both lines showed a significant down-regulation of FaplC, being the percentage of silencing of 84 and 71% on Pel 1 and Pel 3, respectively. The agronomic behaviour of transgenic plants was evaluated during two consecutive years. Fruit set increased in the two transgenic lines when compared with control plants, although Pel 1 showed a significant reduction on fruit weight. Firmness of full ripen fruits from Pel lines was significantly higher than control fruits, while color and soluble solids were not affected. The increase of firmness in Pel lines was maintained when ripe fruits were stored for 3 days at 25 °C. Histological analysis of ripe fruits showed lower intercellular spaces and a higher degree of cell to cell contact area in transgenic fruits when compared with controls. Altogether, these results suggest a direct relationship between pectate lyase gene expression and strawberry fruit softening.     

根癌农杆菌介导蓝猪耳转化系统的建立

 通过对影响根癌农杆菌介导蓝猪耳转化效果各种因素的研究, 建立了蓝猪耳高效稳定的遗传转化系统。研究结果表明, 农杆菌侵染叶片外植体效率最高; 外植体预培养不利于农杆菌的侵染; 乙酰丁香酮能大大提高根癌农杆菌转化蓝猪耳的效率。另外, 菌液浓度、侵染和共培养的时间及再生的途径等均对转化效率有一定的影响。叶片外植体与农杆菌共培养后, 经过抗性芽的诱导、根的再生, 70 d左右就可以获得抗性苗。抗性植株经GUS染色、PCR分析和Southern blot检测, 外源基因已经整合到蓝猪耳基因组中, 转化频率为7%～8%。     

Temperature responses,flowering and fruit yield of the June-bearing strawberry cultivars Florence,Frida and Korona

The effect of night temperature on short day (SD) floral induction has been studied in three June-bearing strawberry cultivars of different geographic origin and compared with yield performance in the cool Nordic environment. At the optimum day temperature of 18 °C, the SD flowering response of the cultivars ‘Florence’ and ‘Korona’ increased significantly with increasing night temperature from 9 to 18 °C, while an optimum was reached at 15 °C in the cultivar ‘Frida’ that is selected under cool-environment conditions in Norway. Also, while saturated flowering response was obtained with 3 weeks of SD treatment at all temperatures in ‘Frida’, several plants of ‘Florence’ and ‘Korona’ failed to initiate flowers at 9 °C night temperature even with 5 weeks of SD. The effect of extended SD period was particularly pronounced in ‘Florence’. The slow SD floral induction response of ‘Florence’ was associated with a 2 week delay of anthesis in subsequent long day (LD) conditions at 21 °C. Yield performance of the same cultivars during 2 years under field conditions at Nes Hedmark and in North Norway also demonstrated that the yield potential of ‘Florence’ was not realized under the climatic conditions prevailing at these locations. In both years the yields varied significantly among the cultivars, ‘Frida’ having the highest yields followed by ‘Korona’, with ‘Florence’ far below. It is concluded that, in the Nordic environment, autumn (September) night temperatures are obviously sub-optimal for yield performance of some June-bearing strawberry cultivars, and that this effect is mediated by autumn temperature effects on flower initiation responses.     

1-MCP对草莓果实采后生理及品质的影响

以转色期丰香草莓为材料,测定了经0.3～0.9μL/L1-MCP处理后果实采后呼吸速率、抗氧化酶活性、活性氧积累以及果实品质的变化规律。结果表明,1-MCP处理抑制草莓果实呼吸作用,维持较高超氧化物歧化酶(SOD)活性和较低的脂氧合酶(LOX)活性,同时降低超氧阴离子(O·2)产生速率、H2O2积累和丙二醛(MDA)含量,有利于保持果实品质。     

农杆菌介导Ferritin基因转化苹果的研究

以皇家嘎拉苹果的叶片为转化受体,通过根癌农杆菌介导将铁结合蛋白(Ferritin)基因导入受体中,诱导获得了抗性芽.经PCR检测从抗Km植株中选择到4株阳性植株,进一步经PCR-Southern杂交证实外源基因已整合到4株苹果基因组中,RT-PCR检测表明,Ferritin基因在4株转基因植株中得到了表达.     

农杆菌介导的百合遗传转化体系的建立

 通过根癌农杆菌介导的遗传转化方法, 建立了凝望星空百合(Lilium ‘Star Gazer’) 愈伤组织和西伯利亚百合(Lilium ‘Siberia’) 叶片的遗传转化体系。试验结果表明: 以愈伤组织和叶片为受体, 均能取得较理想的转化效果; 根癌农杆菌OD600介于0.5～1.0时, 能获得较高的转化率; 40 mg·L ^{- 1}的潮霉素对愈伤组织和叶片有很好的筛选效果; 在共培养培养基中, 去除NH₄NO₃对提高百合转化效率有极显著的效果。对转基因百合进行PCR检测, 结果表明gus基因已整合到百合基因组中。