Saccharum hybrid cultivar (mixed) Transcriptome or Gene expression. Saccharum hybrid cultivar (mixed)

Summary The expression profile of internodes from high brix plant was compared to internodes from low brix plants. Mature (In9), intermediate (In5) and immature internodes (In1) were collected from two different Progenies and immediately frozen in liquid nitrogen. Total RNA was extracted using Trizol. Progeny 1 was derived from two intra-specific polycrosses, one among Saccharum officinarum genotypes and the other combining Saccharum spontaneum genotypes. For each generation, 500 individuals were sampled for brix content and gene expression. The extreme segregants were selected for further analysis. The F3 hybrid individuals selected for molecular studies were planted in a field in single rows of 5 m using standard sugarcane cultivation practices. Brix readings and tissue samples were collected very early in the season, and in March of the following year, when plants were 10 months old. The soluble solids (brix) content of mature internodes of each sugarcane stalk was measured with a portable refractometer (N1 model, ATAGO, Japan). Individuals or pools of eight individuals had their tissues collected and RNA extracted. Progeny 2 was derived from a cross between two commercial varieties (SP80-180 x SP80-4966). Five hundred sugarcane F1 plants were field-grown. The stem sugar content from the different plants showed a normal distribution, and seven plants with extreme brix values were collected. Keywords: Expression profiling by array Overall design: Total RNA from internodes pool from high brix plants and low brix plants was hybridized to dual channel arrays. Internodes from the same stage of development and from the same progeny were compared. The quantification of each hybridization was submitted in two files, one for each slide side (technical replicates).

摘要：本研究对高白利糖度（brix）植株的节间与低糖度植株的节间开展表达谱比较分析。从两个不同的杂交后代群体中分别采集成熟节间（In9）、中间节间（In5）与未成熟节间（In1），样本采集后立即置于液氮中速冻。采用Trizol试剂提取总RNA。
杂交后代1源自两次种内多交：一次为甘蔗（Saccharum officinarum）基因型间的杂交，另一次为野生甘蔗（Saccharum spontaneum）基因型的组合杂交。针对每个后代群体，共采样500个单株用于糖度测定与基因表达分析，选取极端分离单株开展后续研究。用于分子研究的F3杂交单株以5米单行的标准甘蔗栽培模式进行田间种植。于种植季早期以及次年3月（植株生长至10月龄时）采集糖度读数与组织样本。采用便携式折光仪（日本ATAGO公司N1型）测定每根甘蔗茎秆成熟节间的可溶性固形物（白利糖度，brix）含量。采集单株或8株混合的组织样本并提取总RNA。
杂交后代2源自两个商业品种间的杂交（SP80-180 × SP80-4966），共种植500株甘蔗F1代植株。不同植株的茎秆糖含量呈正态分布，最终选取7株具有极端糖度值的单株进行后续分析。
关键词：芯片表达谱分析
整体实验设计：将高糖度植株与低糖度植株的节间混合总RNA进行双通道基因芯片杂交。比较同一发育阶段、同一杂交后代群体的节间样本。每次杂交的定量结果提交为两个文件，分别对应芯片玻片的两个检测通道（技术重复）。