HPLC测定荔枝不同器官中内源激素流程的优化

为了探讨内源激素的提取分离和分析测定的优化流程。试验在前人研究的基础上,以荔枝根、叶、果实为材料,针对荔枝样品富含多酚类和色素等杂质的特点,借鉴酶联免疫法(ELISA)中植物激素提取分离方法,对以往高效液相色谱(HPLC)分析荔枝内源激素的前处理方法进行了改进,配合选择适宜的HPLC色谱条件,摸索出荔枝不同器官中内源激素分析测定的优化流程。结果显示该流程可以联合测定IAA、GA3、ABA和ZT(回收率分别为78.52%、82.36%、79.83%和78.57%),且保留时间稳定,试验重现性好。整个提取分离、分析测定流程简便、快速、准确,适合荔枝材料内源激素的分析测定。     

喷施GA3和2,4-D对留树保鲜脐橙落果和内源激素含量的影响

 以纽荷尔脐橙（Newhall Navel Orange）为试材,测定喷施GA₃和2,4-D的植株留树保鲜果实的落果率和果实内源赤霉素（GA）、生长素（IAA）、玉米素核苷（ZR）、脱落酸（ABA）含量的变化,探讨内源激素与留树保鲜脐橙落果的关系。结果表明：植物生长调节剂处理能降低果实内源GA、IAA、ZR含量下降的速度,减缓内源ABA及ABA/（GA + IAA + ZR）的升高,减少留树保鲜过程中的落果,其中以20 mg · L^-1 GA₃ + 20 mg · L^-1 2,4-D混合处理的效果最好,且留树保鲜应以60 d左右为宜。初步分析认为内源GA、IAA、ZR含量的下降,内源ABA及ABA/（GA + IAA + ZR）的升高加速了果实离层的产生,共同促进了留树保鲜脐橙果实的衰老脱落。     

早蜜梨果实生长发育期间内源激素含量变化的研究

采用高效液相色谱(HPLC)技术,测定了早蜜梨果实生长发育期间内源生长素(IAA)、赤霉素(GA3)、玉米素(ZT)和脱落酸(ABA)含量及其比值的变化,并分析了各激素与果实发育的关系.结果表明:果肉和种子中IAA含量分别在花后50 d和20 d出现高峰;GA3含量分别在花后20 d和60 d达到峰值;ZT含量的高峰则在花后40 d,而ABA含量分别在花后80 d和90 d最高,随后均呈逐渐下降.IAA/ABA、GA3/ABA和ZT/ABA比值对果实的细胞分裂与膨大起着重要调节作用.     

苹果采前落果与内源激素的关系

为了探讨苹果采前落果与内源激素之间的关系,在采前8周间定期测定了不同苹果品种的果柄、果台和离层形成部位组织中IAA、ABA含量;在采收前20d内定期测定了离层部位组织中细胞壁分解酶(Cellulase)的活性;在收获期测定了果实乙烯的发生量。结果表明:不同品种果柄、果台和离层部位组织中IAA和ABA含量变化有差异,但它们变化的总趋势相似,都是随着果实成熟IAA含量下降,而ABA含量上升;不同品种的成熟果实中乙烯发生量有很大差异,以落果多的品种显著大于落果少的品种;采前落果重的品种离层部位组织中细胞壁分解酶的活性在果实成熟期急剧增加。由于果实进入成熟阶段后,IAA含量下降,ABA含量升高,ABA/IAA之间的相对平衡被打破,高ABA/IAA以及高乙烯会刺激离层组织中细胞壁分解酶的活性增高,进而促进离层形成,这可能是导致落果发生的原因。     

梨2个品种果实发育期间内源激素含量的变化

针对不同梨品种果实生长发育历程不同,采用高效液相色谱(HPLC)技术测定了早蜜梨和黄金梨果实发育期间果肉和种子中内源生长素(IAA)、赤霉素(GA3)、脱落酸(ABA)和玉米素(ZT),以及内源腐胺(Put)、亚精胺(Spd)、精胺(Spm)含量的变化。结果表明,1)2品种果肉中IAA、GA3、ABA和ZT含量变化趋势相似,但早蜜梨IAA、GA3含量低于黄金梨,果实发育后期ABA含量高于黄金梨,且提前20d达到含量高峰;2)2品种种子中GA3含量变化分别呈近似单峰曲线和双峰曲线,但早蜜梨IAA、ZT含量高于黄金梨,提前30d达到ABA含量高峰;3)2品种果肉和种子中Put、Spd、Spm含量变化趋势相似,但早蜜梨种子中Put含量低于黄金梨,Spd略高,Spm含量接近。在果实发育前期果肉中Put、Spd含量高于黄金梨。因此,2品种果实发育的各个时期持续时间及其内源激素含量不同。     

架形对“赤霞珠”酿酒葡萄内源激素的影响

以贺兰山东麓酿酒葡萄产区的3个典型小产区直立龙干形和厂字形2种架形种植的“赤霞珠”为试材，采用高效液相色谱法测定了不同生育期功能叶片中的6种植物内源激素，研究了在果实发育关键期架形对叶片内源激素的影响，以期为后期开展不同小产区的品质调控提供参考依据。结果表明：在果实发育关键期，不同处理叶片中的6种内源激素含量差异明显。脱落酸（ABA）、吲哚乙酸（IAA）、反式玉米素核苷（TZR）和赤霉素（GA₃）共同调节酿酒葡萄果实的成熟和品质，成熟期叶片中的吲哚乙酸和单宁呈显著负相关，高浓度的ABA会抑制IAA的合成积累，且转色期ABA的增加直接影响果实中花色苷的合成。不同小产区的生长势和抗性与果实关键发育期的内源激素的比值具有明显的关联性。     

杧果无胚果实内源激素的分离与测定

采用高效液相色谱(HPLC)法测定了杧果无胚果实及正常果实果肉的内源激素(GA3、IAA、ABA和ZT)含量。结果显示:无胚果实果肉的GA3及ZT含量与正常果实无显著差异;无胚果实果肉的IAA含量高达19.91μg·g-1,而正常果实果肉仅为2.44μg·g-1,差异极显著;无胚果实与正常果实的ABA含量分别为3.44μg·g-1和0.023μg·g-1,差异显著。     

两个梨品种果实发育过程中果肉及种子内源激素的变化

为探索梨种子对果实生长发育的影响,通过高效液相色谱(HPLC)技术对"鸭梨"和"新高"果实发育期间果肉和种子中内源生长素吲哚乙酸(IAA)、赤霉素(GA3)、脱落酸(ABA)和玉米素(ZT)含量变化进行了测定。试验结果显示:两个品种种子中ABA和ZT含量变化基本相似,但新高ABA、ZT含量高于鸭梨;鸭梨IAA含量随着果实的发育处于下降趋势,而新高先升高后降低;鸭梨GA3含量随着果实发育先下降后上升,而新高出现两次高峰一次低谷且整体下降的趋势。两个品种果肉中IAA、GA3、ABA和ZT含量变化趋势基本相似,新高IAA、ABA含量高于鸭梨,70d之前新高GA3含量高于鸭梨,然而随着果实发育,鸭梨GA3含量又高于新高,并且鸭梨出现两次高峰,果实发育到80d后,新高ZT含量高于鸭梨。     

根域限制对玫瑰香葡萄果实糖酸及酚类物质和内源激素的影响

[目的]探索根域限制和地栽条件下葡萄果实品质的差异,明确植物内源激素与果实品质形成间的关联,为根域限制栽培技术提高果实品质奠定基础.[方法]以根域限制栽培条件下的玫瑰香葡萄果实为试验材料,测定树体生长指标,采用HPLC和HPLC-MS等方法测定两种栽培模式下果实品质和内源激素的变化,通过相关性分析明确内源激素与果实品质...     

脱落酸和2,4-表油菜素内酯对葡萄成熟过程中果实内源激素含量的影响

以酿酒葡萄"赤霞珠"和"烟73"为试材,研究了脱落酸(ABA)、2,4-表油菜素内酯(EBR)和BR生物合成抑制剂油菜素唑(Brz)处理对葡萄果实乙烯生成量、内源生长素(IAA)和赤酶素(GA3)含量的影响。结果表明:外源ABA和EBR处理促进了果实乙烯的释放,同时促进了果实着色,ABA和EBR处理后果实内源IAA和GA3含量略低于对照,但总体无显著差异。在不同浓度EBR处理中,以0.4mg/LEBR处理效果最显著。Brz处理抑制了乙烯的释放,推迟了果实着色,但对果实内源IAA和GA3含量没有显著影响。     

番茄不同部位中糖含量和相关酶活性的研究

 试验将番茄光合产物运转途径上叶片(源) 、运输系统以及果实(库) 区分开, 分别测定其糖的组成和含量以及糖代谢相关酶的活性。结果表明: 番茄光合产物运转途径上从“源”到“库”各部位糖的组成和含量不同。叶肉中果糖的含量最高, 蔗糖的含量最低; 中筋中以果糖和葡萄糖为主; 叶柄维管束中葡萄糖含量最高, 蔗糖含量次之, 果糖含量最低。节间和果柄维管束中主要含有蔗糖。果实维管束以及果实内各部位中则主要含有葡萄糖和果糖, 且两者含量无显著差异, 蔗糖含量很低。萼片中葡萄糖含量最高, 蔗糖含量最低; 果蒂中3种糖含量均较高且无显著差异。番茄叶肉及光合产物运转组织中转化酶活性很低, 而在库器官的非维管组织中转化酶活性较高。果蒂中的蔗糖合成酶( SS) 活性最高, 其次是叶肉和运转组织, 果实内各部位中SS活性较低。在合成蔗糖的器官—叶肉中, 有较高的蔗糖磷酸合成酶( SPS) 活性, 运转组织中的SPS活性较叶肉中降低, 但果柄维管束和果实维管束中则表现出较高SPS活性, 果肉、果胶质胎座及心室隔壁中的SPS活性最低。     

一种适合于富含多糖和酚类物质的香蕉果实RNA提取方法

酚类化合物和多糖是影响香蕉组织RNA提取的几个主要干扰因素。研究通过在裂解液中加入聚乙烯吡咯烷酮去除多酚类化合物和利用水饱和乙醚去除多糖,改进了香蕉果实RNA的提取方法。该方法具有快速、简单等特点。     

A modified chemiluminescence method for hydrogen peroxide determination in apple fruit tissues

Hydrogen peroxide (H₂O₂) is one of the important by-products produced by plant and fruit tissues during normal metabolism as well as under stress conditions. Evidence suggests that it is actively involved in many physiological activities in plants, including ripening, senescence and the development of disorders. Quantitative measurement of H₂O₂ in fruit has been a challenge due to variations in methodologies, and their sensitivities and interferences present in plant samples. Among the currently used methodologies, chemiluminescence (CL) is one of the most promising, due to its high specificity and sensitivity. However, direct application of CL methods developed for leaf analysis is not suitable for fruit, especially fruit peel tissues, possibly due to interfering compounds in fruit tissues. In this study, evaluation of the efficiency of removal of interfering compounds by PVP, PVPP and activated charcoal revealed that the PVPP is the most effective compound to remove the interference. This modified protocol can measure H₂O₂ content in apple peel and flesh tissues. ‘Red Delicious’ apple peel and flesh tissues were measured with amount of 1.48 and 1.03 μmol/g FW, respectively. The established protocol can also be used for a wide variety of tissues in addition to apple fruit, including strawberry tissues (fruit, calyx and leaves) and spinach leaves. This protocol was applied to determine the H₂O₂ concentration in 1-MCP and DPA treated apples after 5 months of storage, but no significant difference in H₂O₂ in those samples was found. Direct comparison of CL with a commercial hydrogen peroxide measurement OXIS kit was also made. The challenges to accurately assay H₂O₂ in fruit/plant tissue were discussed.     

In vitro culture of albedo tissue from fruits of Citrus sinensis cv. Washington Navel: Effect of fruit age and orange juice

SUMMARY

Callus from citrus fruit explants has been used to study different aspects of fruit physiology. However, growth responses of callus derived from citrus fruit tissues are not always comparable with those of intact fruit; there are, for example, differences in nutritional requirements. Results with tissues cultivated in vitro seem to depend on a variety of environmental and internal factors. In the present work albedo tissue from fruits of Citrus sinensis cv. Washington Navel of different age has been used as a source of explants. The growth (fresh weight) of the calli was dependent on the age of the fruit from which the explants were taken, as well as on the addition of orange juice to the plant growth medium. Orange juice stimulated the growth of the calli and this growth was greater as the fruit age increased. The highest values were obtained when the albedo was isolated from fruits 30 to 45 mm in diameter and grown on medium supplemented with 20% orange juice.     

梨组织RNA提取方法研究

借鉴荔枝果实总RNA提取方法,针对梨本身的特点利用改良 Bugos法分别对叶片、花瓣、果肉中总RNA进行提取研究.结果表明:改良Bugos法对梨不同组织中RNA的提取效果较好,花瓣中RNA的相对含量最高,叶片次之,果肉最少.     

Cultivar-based fruit size in olive depends on different tissue and cellular processes throughout growth

In drupe fruits, in addition to fruit size, the proportions of mesocarp and endocarp tissues are critical objectives for fruit quality, crop production and management. The olive fruit is a typical drupe, with cultivars which show a wide range in both fruit size and the proportions of mesocarp and endocarp. Characterizing the roles of tissue and cellular processes in producing genetically based fruit size variability is necessary for crop improvement, as well as deepening our understanding of fruit developmental physiology. This study used microscope image analysis to evaluate cell number and size, the growth of mesocarp and endocarp tissues, and their developmental timing in producing fruit size among six olive cultivars with a large range of fruit size. We found that cultivar mesocarp and endocarp size increased linearly with fruit size, with larger sizes favoring an increasingly greater mesocarp/endocarp ratio. Within the mesocarp, cultivar-based fruit size related directly to cell number and was established soon after bloom by cell division rate. In spite of different cell division rates, all cultivars showed similar timing of cell division activity, with the majority of cells produced in the two months after bloom but, surprisingly, a substantial number of cells formed during the following 6 months. Cell expansion was high throughout fruit growth and an important factor in achieving final fruit size, but cell size did not differ among cultivars at any time. We can conclude that fruit size differences among olive cultivars are due at the tissue level to both mesocarp and endocarp sizes and at the cellular level to cell division throughout fruit growth. Furthermore, since cell size is consistent among cultivars in spite of variable cell division, it is likely that cultivar differences in cell expansion accompany those in cell division.     

野生欧李生长期组织器官中不同形态钙含量的变化及其相关性

 研究了野生欧李萌动期、开花期和果实发育期各组织器官中不同形态钙含量的变化及其相关性。结果表明,野生欧李对钙的吸收积累与钙的形态密切相关,木质部和花器官、果实、果柄中的钙主要以果胶钙形态为主,韧皮部和新梢中的钙主要以草酸钙形态为主。在年生长发育时期,木质部中各种形态钙含量总体呈下降趋势,进入果实成熟膨大期含量回升;韧皮部中各种形态钙含量总体呈下降趋势,进入果实成熟膨大期水溶钙、果胶钙、磷酸钙和草酸钙含量略有回升;花器官和果实中各种形态钙含量变化呈“下降—增加（或下降）—下降”的变化;在果实发育期,新梢中的水溶钙、果胶钙、草酸钙和剩余钙含量明显增加,而果柄中的水溶钙含量增加最为明显。随生长发育进程各组织器官中存在钙形态转化。木质部、韧皮部、花和果实中水溶钙和果胶钙含量的年变化与相应器官中钙总量之间存在高度正相关。     

佛手果形发育观察及果形相关基因表达分析

通过超景深显微镜和扫描电镜观察不同发育阶段的佛手（Citrus medica L. var. sarcodactylisSwingle）花芽形态,发现佛手指状部分并非发育自柱头,而是来自雌蕊子房。同时,在佛手中分离了CmsSUN20、CmsOFP7、CmsOFP12 和CmsYABBY5 等4 个果形相关基因的全长,与甜橙序列比对的相似性大于95%,构建进化树分析发现其与甜橙聚在一个分支。收集佛手和香橼（C. medica L.）果实发育相同阶段不同组织（叶片,花瓣,雌蕊,雄蕊）材料,以qRT-PCR 法分析了上述4 个基因的表达。结果表明CmsSUN20 在佛手雌蕊的表达显著高于香橼,在其他组织中差异不显著或相反,表明该基因可能与佛手雌蕊发育有关。CmsYABBY5 在佛手雌蕊和雄蕊中都显著高表达,而在叶片中却显著低表达,表明CmsYABBY5 有可能对佛手雌蕊和雄蕊发育有重要作用。而CmsOFP7 和CmsOFP12 在佛手叶片、雌蕊和雄蕊中均高表达,由于并非特异在雌蕊中高表达,所以可能并不对佛手果形建成起决定作用。     

荔枝果柄结构发育与落果的关系

在荔枝种子发育的各阶段，对落果与坐果果柄进行比较解剖观察，结果表明，落果果柄各组织、细胞的数量和分化速度均不及坐果果柄。花期和胚乳核分裂前期，花及幼果脱落与其果柄初生维管束和导管数量较少有关。在合子分裂期，果柄次生输导组织和中柱鞘纤维的产生，以及胚体形成期皮层和中柱鞘分化的石细胞，均可加强果柄机械力度。果熟期由于果柄周皮形成和内部结构完善，落果数量减少。