【文献标题】 Cytological, physiological, ａnd transcriptomic analyses of golden leaf coloration in Ginkgo biloba L

【作者】 Wei-xing Li, Shun-bo Yang, Zhaogeng Lu，et.al

【作者单位】 扬州大学

【文献中引用产品】

有机氯elisa试剂盒（有机氯农药残留快速检测ELISA检测试剂盒）

【DOI】https://doi.org/10.1038/s41438-018-0015-4

【影响因子(IF)】 4.21

【出版期刊】 《Horticulture Research》

【产品原文引用】

Materials ａnd methods

Plant material

A healthy adult G. biloba tree bearing both normal green leaves ａnd golden–green striped leaves was used in this study, after growing under natural conditions in the Ginkgo nursery of Yangzhou University (32°39′ N, 119°42′E). Normal green leaves ａnd golden–green striped leaves borne on the same short shoot of this tree were collected from May to July in 2015 ａnd 2016 (Fig. 1a–c). For cytological, physiological ａnd RNA-Seq experiments,yellow parts of the golden–green striped leaves (mutant leaves) ａnd the green leaves (normal leaves) were sampled separately in May. For RT-qPCR, the mutant leaves ａnd green leaves from three stages (May to July) were used. All of the samples were collected from the identical spatial sections of leaves (Fig. 1d, e), ａnd immediately frozen in liquid nitrogen, ａnd stored at −80 °C until use.

Measurements of chlorophyll, carotenoid, ａnd flavonoid contents  Approximately 0.1 g of leaves from normal green leaves ａnd the golden parts of mutant leaves (sampled in May) were cut into pieces and  submerged in 80% acetone Fig. 1 Phenotypes of normal green leaves ａnd mutant leaves from the same short shoot of Ginkgo biloba in May. a Phenotype of the G.biloba tree. b, d Phenotype of the normal green leaf. c, e Phenotype of the mutant leaf. The red-dashed circles near arrows represent the sampling areas Li et al. Horticulture Research (2018) 5:12 Page 2 of 14 overnight to extract the chlorophyll. Next, the extract was measured spectrophotometrically at 665, 649, ａnd 470 nm with a UV-1800 spectrophotometer (DU-800; Beckman Coulter, Brea, CA, USA). Soil-plant analysis development (SPAD) values were measured in situ on each type of leaf using a Chlorophyll Meter SPAD-502Plus (Konica Minolta, Osaka, Japan). To measure the contents of chlorophyll intermediaries, leaves were homogenized in nine volumes of 0.01 M phosphate-buffered saline, mixed on ice, ａnd centrifuged (30 min at 2500 g). The supernatant was assayed separately with an ELISA kit (HengYuan Biological Technology Co., Ltd, Shanghai, China). Carotenoid components were measured using highperformance liquid chromatography, ａnd total carotenoid ａnd flavonoid contents were determined using a UV-1800 spectrophotometer. Three biological replicates were evaluated for each sample. The data thus obtained were analyzed using SPSS software (ver. 17.0; SPSS Inc.,Chicago, IL, USA)

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