共查询到20条相似文献，搜索用时 15 毫秒
1.
【目的】探究在不同成熟度和降温贮藏方式下,LAC表达模式与鸭梨果心褐变的关系,为进一步解析鸭梨果心褐变机制提供理论依据。【方法】以鸭梨作为材料,通过对不同成熟度（早采、中采和晚采）的鸭梨进行急速降温（急降）和缓慢降温（缓降）处理,观察贮藏期间鸭梨果心褐变情况,测定漆酶（Laccase,LAC）活性及其基因LAC的相对表达量,研究LAC在鸭梨果心褐变过程中的参与作用。【结果】冷藏60 d时,晚采鸭梨出现褐变,晚采缓降处理的鸭梨果心褐变指数为0.32,是同期急速降温处理的2.56倍;在贮藏90 d时,中采缓降处理的褐变指数是0.24,中采急降处理的褐变指数仅为0.01。各个处理组在贮藏期间LAC活性多数表现为先逐渐升高后下降的趋势,晚采的果实在贮藏60 d时出现活性高峰,褐变发生;早采和中采鸭梨LAC活性高峰均在90 d时出现,褐变程度低于晚采鸭梨。在贮藏期间,缓慢降温处理的LAC活性高于急速降温处理。鸭梨LAC14和LAC7表达量呈先上升后下降的趋势,LAC6表现为先下降后上升再降低的变化趋势;中采、晚采鸭梨在贮藏60 d时,LAC14和LAC7表达量最高。【结论】与缓慢降温相比,急速降温处理减少了鸭梨果心褐变的发生。在整个贮藏期间,LAC活性呈先上升后下降然后又上升的趋势,其峰值时的活性高低次序为：晚采>中采>早采,这与果心褐变趋势一致;LAC在鸭梨果心褐变过程中上调表达。相比缓慢降温处理,急速降温处理具有较低的LAC7、LAC14和LAC6表达量。急速降温结合适时采收能够抑制LAC的上调表达,减少鸭梨果心褐变发生。 相似文献
2.
Studies on browning problem and phenols content on shoots of Yali, Aikansui and Abbe Fetel pears for in vitro culture 总被引：1，自引：0，他引：1
Bharat Kumar Poudyal Guoqiang Du Yuxing Zhang Jie Liu Qingchun Shi 《Frontiers of Agriculture in China》2008,2(3):321-330
Yali is one of the best pear cultivars cultivated extensively in China and other countries. However, mortality rates of explants
during the initial phase of tissue culture were found to be very high during the summer particularly from the shoot tip explants.
A thorough investigation on the browning problem of Yali Aikansui and Abbe Fetel pears was done and their control measures
are suggested in this paper. Emphasis has been given to the Yali pear. Shoot explants were collected from the field as well
as from the plantlets grown in vitro during different months and observed for browning. The explants were excised and cultured in Murashige and Skoog (MS) solid
and liquid medium as needed. The cultures were maintained in the dark as well as in a 16/8 hours light/dark photoperiod regime
as needed and were kept in a growth room at (25±2)°C temperature. According to the experimental results, Yali was found more
severely infected by browning than the other two cultivars. Similarly, it was also found that, the intensity of browning was
less during spring and increased with time and reached the maximum during summer. Shoot tips of Yali were found more severely
infected by browning than the second node and other nodes. This accelerated the mortality rate up to 81% of shoot explants
during the summer months of July and August. Browning was greatly influenced by water-soluble polyphenols, more of which were
found in the shoot tip of explants collected from the field (significantly higher at P ⩽ 0.05 and P ⩽ 0.01). Total polyphenol contents were found less in the explants from plantlets grown in vitro, and less browning of medium
appeared resulting in a lower mortality rate of explants. Collections of explants during early spring (P ⩽ 0.05 and P ⩽ 0.01) and use of other nodes rather shoot tip because explants (P ⩽ 0.05 and P ⩽ 0.01) were found to be better in preventing the browning problem. Finally, for curative measures to control browning, use
of ascorbic acid at the rate of 100 mg·L−1 (P ⩽ 0.05 and P ⩽ 0.01), 0.02%polyvinyl pyrrolidone (PVP) in the culture medium (P ⩽ 0.05), 96 hours dark treatment of other nodes (P ⩽ 0.05), and 12 hours cold treatments of explants at 4°C (P ⩽ 0.05 and P ⩽ 0.01) prior to sterilization of explants, were found to be the best methods to control browning and therefore to increase
the survival rate of cultured explants of the Yali pear. 相似文献
3.
【目的】从鸭梨中克隆GAPDH基因家族,筛选合适的鸭梨内参基因。【方法】采用同源克隆的方法,克隆鸭梨GAPDH基因家族,用半定量PCR方法分析4个鸭梨GAPDH基因在鸭梨幼叶、花蕾、盛花期花瓣、幼果期果肉和成熟期果心,以及幼果期、膨大期、成熟期和褐变期果心的表达特征。【结果】从鸭梨中成功克隆出了4个GAPDH基因,分别命名为PbGAPDHa、PbGAPDHb、PbGAPDHc1和PbGAPDHc2。其中PbGAPDHa和PbGAPDHb是由一个共同的祖先基因倍增进化来的,主要在鸭梨的幼叶中表达,其蛋白定位在质体;而PbGAPDHc1和PbGAPDHc2是一个基因的2种拷贝形式,在鸭梨的花、幼叶、果实等组织中表达量基本一致,在不同发育时期的果心中表达量也相近,其蛋白定位在细胞质。【结论】PbGAPDHc1和PbGAPDHc2适合作为鸭梨的内参基因。 相似文献
4.
To investigate the inductive effect of salicylic acid (SA) on the resistance of Pyrus bretschneider cv Yali to black spot disease (Alternaria kikuchiana Tanaka), the physiological and biochemical characteristics of detached pear leaves at the age of 5 to 10 days were measured
after application of SA. The results showed that exogenous SA significantly improved the resistance of Yali pear (Pyrus bretschneider cv Yali) leaves to black spot disease. For the SA treatment at 0.02 mmol·L−1 SA concentration, the disease index was the lowest, and the induced resistance reached up to 63.9%. Furthermore, SA induced
local and systemic resistance of Yali pear against the black spot disease. Expression of systemic resistance in leaves was
detectable 3 d after SA treatments and lasted for 10 d. POD, PPO, and PAL activities of Yali pear leaves increased by SA treatment.
It is suggested that exogenous SA solution as a chemical activator could induce the resistance of Yali pear to black spot
disease. 相似文献
5.
水稻OsCER4基因编码脂肪醛脱羧酶,与拟南芥CER1基因高度同源,是否参与植物表面蜡质合成尚不清楚,为了探讨其功能,扩增了OsCER4基因起始密码子ATG上游约2 kb的序列作为该基因的启动子,以常规技术将启动子和OsCER4基因反义片段克隆到pCAMBIA双元载体1380中,采用农杆菌介导法转化粳稻品种中花11,并进行了转化苗OsCER4蛋白的表达量变化分析.结果表明,成功构建了由OsCER4自身启动子驱动的反义RNA载体,并通过农杆菌介导法成功转入水稻中花11中,多数OsCER4基因反义转化植株为阳性转化植株,后代分离比符合3∶1,反义RNA转化植株在蛋白水平上的表达量下降. 相似文献
6.
自花结实鸭梨品种自交亲和分子机理研究 总被引：1，自引：0，他引：1
对鸭梨及其自花结实变异品种-金坠梨和魏县阎庄自花结实鸭梨的S基因进行了研究。通过对3个品种基因组DNA进行S基因特异性PCR扩增,扩增产物经2%琼脂糖凝胶电泳检测发现,在370 bp左右均扩增出了1条带,但不能区分2个S等位基因;经8%聚丙烯酰胺凝胶电泳检测发现,鸭梨和金坠梨在370 bp左右有2条带,而魏县阎庄自花结实鸭梨只有1条带,表明魏县阎庄自花结实鸭梨系鸭梨的1个S基因发生了变异,从而导致自交亲和;进一步研究表明,魏县阎庄自花结实鸭梨S1基因正常,系S21基因发生了变异,导致自交亲和;金坠梨和鸭梨谱带无差别,自交亲和机理可能是调控S基因表达的修饰基因发生了变异,导致自交亲和。 相似文献
7.
The study analyzed the silencing of BcMF12 gene regulated by BcA9 promoter in the transgenic pakchoi and confirmed the effect of antisense BcMF12 gene on the pollen development. A conserved BcMF12 gene fragment was amplified from the cDNA of flower buds in pakchoi （Brassica campestris L. ssp. chinensis, syn. B. rapa L. ssp. chinensis） and was fused to the anther specific BcA9 promoter. The plant antisense expression vector was constructed and then introduced into pakchoi via Agrobacterium-mediated transformation. The transgenic plants were screened by antibiotics and molecular analysis. PCR and Southern blot revealed that the antisense BcMF12-GUS fusion gene regulated by BcA9 promoter was integrated into transgenic plants. Northern blot suggested that the expression of BcMF12 gene was down-regulated significantly. The pollen germination rate of transgenic plants with antisense BcMF12 gene decreased as compared with that of the control plants. The expression of the gene BcMF12 related to the pollen development was inhibited by the antisense BcMF12 driven by BcA9 promoter, which consequently affected the pollen development in pakchoi. 相似文献
8.
为揭示梨果实石细胞分化的机理,调控石细胞形成和含量,克隆了鸭梨(Pyrus bretschneideri Rehd.‘Yali’)4-香豆酸∶辅酶A连接酶基因Pb4CL片段(Gen Bank登录号:KF177692),该片段长467 bp,经同源性比对,该片段与沙梨、欧洲花楸和枇杷等物种4CL基因高度同源,编码155个氨基酸残基,存在4CL蛋白质高度保守区域,具有腺苷酸合成class I超级结构域。建立了梨Pb4CL基因的荧光实时定量表达检测体系,在石细胞分化期Pb4CL基因相对表达量逐渐升高,后呈下降趋势。 相似文献
9.
为了了解鸭梨采后种子生理及转色与鸭梨果实褐变的关系,研究了降温方法对不同采收成熟度鸭梨种子转色和细胞膜透性的影响。结果表明:随着采收成熟度的提高和贮藏期的延长,鸭梨种子颜色逐渐加深,转色指数增加;采后早采鸭梨种子PPO活性和细胞膜渗透性最高,中采果次之,晚采果最低,较高的PPO活性导致早采果种子颜色变化最大;缓慢降温较好地抑制了鸭梨种子转色指数、PPO活性和细胞膜渗透率的升高,而且对早采果种子的影响更为明显。 相似文献
10.
以苹果梨果皮为试材,克隆了苹果梨果实着色相关PAL基因片段,GenBank登录号为JN120855。结果表明:该基因片段长度为407bp,与鸭梨PAL基因序列一致性达97%,与鸭梨氨基酸序列同源性最高,酶切位点分析表明,该PAL基因序列含有常用的限制性内切酶AccI的识别位点。半定量RT-PCR分析表明,随着果实的成熟,套袋果与未套袋果PAL基因的表达量都不断增加,但套袋果在果实成熟期的表达量明显高于未套袋果。 相似文献
11.
【目的】筛选与FWL1互作的蛋白,为研究FWL1基因调节果实大小机理提供依据。【方法】以杜梨、库尔勒香梨、鸭梨、早美香为材料,提取4个梨品种果实细胞分裂关键时期的果肉组织混样RNA。构建梨幼果FWL1膜系统酵母双杂交三框cDNA文库,并鉴定文库质量。构建诱饵载体并转化酵母菌,筛选与FWL1互作的蛋白,对初步筛选出的阳性酵母克隆进行测序并在NCBI中进行Blast比对,确定候选互作蛋白。【结果】文库库容约为3×107 CFU,大于1×107 CFU,平均插入片段大于1 000 bp,阳性率为≥98%。诱饵载体无自激活功能,利用共转化方法,筛选出272个与FWL1互作的蛋白质。【结论】梨幼果膜系统酵母双杂交三框cDNA文库符合酵母双杂文库的基本条件,适用于后期筛选相互作用的蛋白。筛选出的272个互作蛋白在果实发育过程中表达不同,其中collagen and calcium-binding EGF domain-containing protein 1和metallothionein-like protein可能与梨果实大小发育有关。 相似文献
12.
缺钙会引起许多果实生理性病害。为给梨树合理补钙提供理论依据,以7个梨主栽品种为试材,研究了其叶片、果实和新梢钙含量的动态变化规律。结果表明：随着梨的生长发育,参试品种的叶片和新梢钙含量均大体呈增加趋势,果实钙含量呈逐渐降低趋势（其中幼果期至果实成熟前期降速最快）,且同一品种幼嫩器官与成熟器官的钙含量差异均达到了显著水平,同一发育时期相同器官钙含量最大值与最小值的差异也达到了显著水平。钙含量最高的器官是叶片,其次是新梢,果实钙含量最低。不同梨品种的钙素营养水平存在差异,其中各器官发育末期的钙含量,均以鸭梨最高,黄冠和大南果较低。 相似文献
13.
LIU Le-cheng CAO Jia-shu YU Xiao-lin XIANG Xun FEI Yong-jun 《中国农业科学(英文版)》2006,5(5):339-345
In an effort to provide some information relevant to the molecular mechanism of genic male sterility in plants, BcMF3 gene that encodes a pectin methylesterase was isolated from the fertile B line of Chinese cabbage-pak-choi （Brassica rapa ssp. chinensis, syn. B. campestris ssp. chinensis）. In the present paper, a 455-bp antisense cDNA fragment of BcMF3 was introduced to binary vector pB1121, and then was mobilized into Agrobacterium tumefaciens strain LBA4404. The A. tumefaciens harboring the BcMF3 antisense fragment was transformed to Arabidopsis thaliana by floral dip. Scanning electronic microscopy examination demonstrated that 47.8% of BcMF3 antisense pollen grains exhibited abnormal shape, which might lead to decreased germination of pollens, suggesting that the product of BcMF3 gene plays an important role during microsporogenesis. The evidence on burst of 45.7% of BcMF3 antisense pollen tubes in vitro and a majority of BcMF3 antisense pollens restricted within the stigmatic tissue revealed that BcMF3 is involved in aiding the growth of pollen tubes. The results suggest that BcMF3 acts at both stages of microsporogensis and pollen tube growth. 相似文献
14.
Chunli Song Junlian Ma Xia Tang Zide Zhang Zhixia Hou 《Frontiers of Agriculture in China》2009,3(4):419-424
The short hairpin RNA (shRNA) expression vector of the FaEtr2 gene was constructed by inserting the sense fragment into the constructed antisense vector of FaEtr2 (pBI121-Anti-Etr2) in sense orientation. The constructed RNA interference (RNAi) expression vector was transformed into Agrobacterium fumefeciens LBA4404 and used to infect strawberry leaves. Using in vitro plantlet leaves as explants, the transformation conditions of All-Star strawberry were studied systemically. The results
showed that infecting the leaves with A. fumefeciens resuspension liquid of OD600 = 1.0 by pre-culturing for 3 d, co-culturing for 3 d, infecting for 10 min, and adding acetosyringone (AS) 50–100 μmol·L−1 was suitable for genetic transformation of All-Star strawberry. Seven lines of transgenic plants were preliminarily identified
by PCR and β-glucurondiase (GUS) histochemical staining. 相似文献
15.
将水稻DAD1基因插入Ti质粒,构建成该基因的正反义植物表达载体pWM-DAD1,pWM-antisense DAD1,转入根癌农杆菌LBA4404后,利用农杆菌介导的叶盘法转化菊花,经Hyg抗性筛选,并诱导愈伤组织和分化成苗,得到转基因菊花植株,通过PCR检测,从部分转基因植株中检测出预期的目的基因的存在. 相似文献
16.
油菜PEP基因的克隆及PEP反义基因的构建 总被引：17，自引：0，他引：17
丙酮酸羧化酶（ Ｐ Ｅ Ｐ）是控制油菜蛋白质／油脂含量比例的一个关键酶⒚本研究用 Ｐ Ｃ Ｒ 法扩增出了 Ｐ Ｅ Ｐ 基因片段,并将其 克隆到 ｐ Ｂ Ｓ Ｓ Ｋ＋ 的 Ｓｍ ａ Ⅰ位点⒚ Ｄ Ｎ Ａ 序列分析表 明克隆片段 的长度为 ５３０ｂｐ,其序列与报道序列相同⒚将该 Ｐ Ｅ Ｐ基因 片段反向插入 ｐ Ｉ Ｇ１２１ 质粒,构建了带 Ｐ Ｅ Ｐ 反义基因的超级双元载体并进行油菜的转化,目前已获得转基因植株⒚ 相似文献
17.
自花结实鸭梨授粉受精过程中生理代谢变化规律研究 总被引：1，自引：0，他引：1
对鸭梨及其自花结实变异品种--"金坠梨"和"魏县阎庄自花结实鸭梨"的自花授粉表现、花柱抗氧化酶活性、MDA含量、内源激素含量及比值进行了研究.结果表明:"金坠梨"和"魏县阎庄自花结实鸭梨"自花授粉后,花粉管能进入花柱,在沿花柱向子房伸长途中未被抑制,花粉管能顺利通过花柱进入胚囊,并能完成受精作用.鸭梨的自花结实变异品种... 相似文献
18.
牡丹ACC氧化酶基因片段的克隆及其反义表达载体的构建 总被引：1，自引：1，他引：0
利用CTAB改良法提取牡丹洛阳红花瓣总RNA,通过反转录酶和设计的ACC氧化酶基因特异引物合成709bp目的片段,进行测序和比对,以确定目的片段的正确性。在709bpACC氧化酶基因片段和pBI121植物表达载体上选择合适的酶切位点,进行XbaⅠ和SmaⅠ双酶切。将709bp片段上切下的510bp片段反向插入pBI121植物表达载体35S启动子后面,构建成含510bp牡丹ACC氧化酶基因反义表达载体。最后采用电击法转化进感受态农杆菌GV3101,以备用于花粉管通道法转化牡丹。 相似文献
19.
[目的]为研究水稻中的SLR1蛋白的功能及寻找与其相互作用的GA信号转导蛋白奠定基础。[方法]以水稻品种日本晴基因组DNA为模板,根据水稻SLR1基因cDNA序列设计1对特异引物进行PCR扩增,回收PCR产物连接到pGEMT载体中,经筛选得到水稻SLR1基因的克隆pGEMTSLR。最后采用酚仿抽提和乙醇沉淀的方法构建水稻SLR1基因的正义和反义表达载体。[结果]通过PCR扩增获得了约为1.9kb的特异片段。回收PCR产物,将克隆片段和pGEMT载体连接后转化到大肠杆菌DH5α感受态细胞,重组克隆经酶切鉴定后提取重组质粒pGEMTSLR1。该研究成功构建了SLR1基因的正义表达载体pCAMSLR(约0.7kb)和反义表达载体pCAMASLR(约1.2kb)。[结论]采用冻溶法可将表达载体pCAMSLR和pCAMASLR导入农杆菌并进行水稻的遗传转化。利用SLR1基因的正义和反义表达载体,可观察转基因植株中该基因的表达上调和下调对植物的影响。 相似文献