SlMYB94轉殖基因對番茄花器長度的影響評估

Assessing Effect of the Transgenic SlMYB94 Allele on Floral Organ Length in Tomato

陳品堯(Pin-Yao Chen)

指導教授 ： 陳凱儀

國立臺灣大學/生物資源暨農學院/農藝學研究所/碩士(2017年)

https://doi.org/10.6342/NTU201704293

摘要

番茄的se2.1數量性狀基因座為複合式數量性狀基因座，包含style2.1、stamen2.1、stamen2.2、stamen2.3、以及dehiscence2.1，其中的stamen2.2的候選基因定位在番茄第二對染色體上cLED19A24至CT9分子標記區間內。本研究選擇位於此染色體區間內的SlMYB94基因為stamen2.2的候選基因，並藉由基因轉殖的方式，觀察SlMYB94對偶基因數目的增加，是否造成番茄雄蕊與雌蕊長度的改變。轉殖植株的初步篩選是採用CaMV 35S分子標記和番茄背景分子標記對轉殖T_0世代植株進行篩選，T_1自交世代個別植株的基因型則藉由焦磷酸定序 (pyrosequencing) 所偵測轉殖對偶基因型與背景對偶基因型之單核苷多型性的比例推算而得。試驗結果顯示T_0世代轉殖株「中研院2-9」成功轉殖單一SlMYB94對偶基因，然而在其自交世代16TS357族群中，帶有不同數目轉殖對偶基因植株之間的雄蕊與雌蕊長度並沒有顯著的差異。由此結果推論SlMYB94並非stamen2.2基因。

關鍵字

番茄 ； se2.1 ； stamen2.2 ； MYB ； 焦磷酸定序 ； 雄蕊長

並列摘要

The tomato se2.1 is a composite quantitative trait locus, including style2.1, stamen2.1, stamen2.2, stamen2.3, and dehiscence2.1. The stamen2.2 was delimited between molecular markers cLED19A24 and CT9 on the chromosome 2. In the current study, the SlMYB94 gene is one of genes residing in the aforementioned chromosomal region and was chosen as the candidate gene of stamen2.2. The correlation between the SlMYB94 copy number and the changes of stamen and style length were observed via the gene transformation approach. CaMV 35S marker and genetic background marker were used to identify successful transgenic plants in the T_0 generation. In the T_1 generation, pyrosequencing was used to detect the proportion of single nucleotide polymorphism between the transformed allele and the background allele, and hence to infer genotypes of T_1 plants. The results showed that the “sinica2-9” T_0 plant successfully received a single copy of the SlMYB94 transgenic allele but its T_1 progenies, the 16TS357 population, did not show significant difference on style length and stamen length between individuals carrying different copy number of the SlMYB94 transgenic alleles. This conclusion infers that the SlMYB94 gene is not stamen2.2.

並列關鍵字

tomato ； se2.1 ； stamen2.2 ； MYB ； pyrosequencing ； stamen length

參考文獻

Chen KY, Tanksley SD. (2004) High-Resolution Mapping and Functional Analysis of se2.1: A Major Stigma Exsertion Quantitative Trait Locus Associated With the Evolution From Allogamy to Autogamy in the Genus Lycopersicon. Genetics 168: 1563–1573.

Daminato M, Masiero S, Resentini F, Lovisetto A, Casadoro G. (2014) Characterization of TM8, a MADS-box gene expressed in tomato flowers. BMC Plant Biology 14:319

Dobritzsch S, Weyhe M, Schubert R, Dindas J, Hause G, Kopka J, Hause B (2015) Dissection of jasmonate functions in tomato stamen development by transcriptome and metabolome analyses. BMC Biology 13:28

Gates DJ, Strickler SR, Mueller LA, Olson BJSC, Smith SD. (2016) Diversification of R2R3-MYB Transcription Factors in the Tomato Family Solanaceae. Journal of Molecular Evolution 83:26–37

Kiferle C, Fantini E, Bassolino L, Povero G, Spelt C, Buti S, Giuliano G, Quattrocchio F, Koes R, Perata P, Gonzali S. (2015) Tomato R2R3-MYB Proteins SlANT1 and SlAN2: Same Protein Activity, Different Roles. PLoS ONE 10(8): e0136365.