利用農桿菌滲入法與農桿菌注射法建立監測植體含磷狀態之系統

Development of an in Planta System to Monitor Phosphorus Status by Agroinfiltration and Agroinjection

林佳螢(Jia-Ying Lin)

指導教授 ： 林淑怡

共同指導教授 ： 羅筱鳳(Hsiao-Feng Lo)

國立臺灣大學/生物資源暨農學院/園藝暨景觀學系/碩士(2014年)

https://doi.org/10.6342/NTU.2014.00991

摘要

磷 (Phosphorous, P) 是植物生長發育所需必要營養元素之一，參與調控植物體內多項生理、生化反應。本研究於菸草與番茄之營養生長期進行缺磷處理，地上部與根部之無機磷濃度快速降低，葉綠素螢光Fv/Fm讀值、葉綠素相對含量與地上部鮮重的減少較緩慢，而根部對地上部鮮重之比值則逐漸增加。再者，番茄植株於生殖生長期遭遇缺磷逆境時，果實之無機磷濃度快速降低，進而降低果實產量與可溶性固形物含量。以上結果說明菸草與番茄植株的營養生長以及番茄果實最終之產量與品質皆仰賴於磷肥的充足供應。本研究進一步發現，短暫缺磷雖然會迅速降低地上部或果實的無機磷濃度，但是若在尚未對植株地上部鮮重或果實產量與品質造成不利影響前，迅速補充磷肥，可迅速提升地上部或果實中的無機磷濃度，因而不會對植株地上部鮮重或果實的產量與品質造成不利影響。這說明透過監測作物營養狀態，而適當的供給肥料將可確保作物的產量與品質。本研究透過農桿菌滲入法和農桿菌注射法的方式，證實GUS報導基因在番茄TPSI1啟動子的驅動下，可快速、忠實反應菸草葉片與番茄果實面臨缺磷逆境的情形，故知農桿菌滲入法和農桿菌注射法短暫表現的系統可用以監測植體缺磷狀態。而 GUS報導基因的表現並不受磷以外的其他營養元素缺乏所影響，說明此套農桿菌滲入法短暫表現的系統具專一性。由於此套系統可方便、快速、準確反應植體缺磷狀態，未來將可進一步應用於其他作物；若透過以其他適合的啟動子取代TPSI1，此套系統也可應用於監測其他營養元素之狀態。

關鍵字

磷 ； 菸草 ； 番茄 ； 農桿菌滲入法 ； 農桿菌注射法

並列摘要

Phosphorous (P), one of the essential mineral nutrients for plant growth and development, is involved in the regulation of several physiological and biochemical processes in plants. In our study, it was found that, in the tobacco and tomato plants at vegetative growth stage under phosphorus deficiency treatment, the phosphate (Pi) concentration in shoot and root decreased rapidly, the chlorophyll fluorescence (Fv/Fm), chlorophyll content and shoot fresh weight decreased relatively slowly, while the root/shoot fresh weight ratio increased gradually. Furthermore, in tomato plants at reproductive growth stage under Pi starvation, the Pi concentration in fruit decreased rapidly, and then the fruit yield and total soluble solids decreased. Accordingly, it is shown that the vegetative growth of tobacco and tomato plants as well as the final yield and quality of tomato depend on whether phosphorus fertilizer is supplied sufficiently. It was further found that, while temporary phosphorus deficiency may immediately lead to the decrease of the Pi concentration in shoot or fruit, the shoot fresh weight or the fruit yield and quality may not be adversely impacted thereby so long as phosphorus fertilizer is timely supplemented to rapidly increase the Pi concentration in shoot or fruit. This indicates that the yield and quality of crops can be ensured by monitoring the nutritional status of the crops and supplying fertilizer as appropriate. By Agroinfiltration and Agroinjection, this study demonstrated that the expression of GUS reporter gene driven by tomato TPSI1 promoter can rapidly and truly reflect the phosphorus deficiency stress in tobacco leaf and tomato fruit. This indicates that the employed Agroinfiltration/Agroinjection transient expression system is useful in monitoring the Pi status in plants. Further, the aforesaid expression of GUS reporter gene is independent of the deficiency of the mineral nutrients other than phosphorus, indicating the specificity of the employed Agroinfiltration transient expression system. Because the application of the aforesaid system is convenient and leads to rapid and accurate reflection of phosphorus status in plants, the aforesaid system should be applicable to other crops or to monitor other mineral nutrient status if the tomato TPSI1 promoter is replaced by other appropriate promoters.

並列關鍵字

phosphorus ； tobacco ； tomato ； agroinfiltration ； agroinjection

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