[논문]Tomato Yellow Leaf Curl China Virus Impairs Photosynthesis in the Infected Nicotiana benthamiana with βC1 as an Aggravating Factor

Farooq, Tahir; Liu, Dandan; Zhou, Xueping; Yang, Qiuying

doi:10.5423/ppj.oa.04.2019.0120

Tomato Yellow Leaf Curl China Virus Impairs Photosynthesis in the Infected Nicotiana benthamiana with βC1 as an Aggravating Factor 원문보기

The plant pathology journal, v.35 no.5, 2019년, pp.521 - 529

Farooq, Tahir (State Key Laboratory for Plant Disease and Insect Pest, Institute of Plant Protection, China Academy of Agricultural Sciences) , Liu, Dandan (State Key Laboratory for Plant Disease and Insect Pest, Institute of Plant Protection, China Academy of Agricultural Sciences) , Zhou, Xueping (State Key Laboratory for Plant Disease and Insect Pest, Institute of Plant Protection, China Academy of Agricultural Sciences) , Yang, Qiuying (State Key Laboratory for Plant Disease and Insect Pest, Institute of Plant Protection, China Academy of Agricultural Sciences)

Abstract ▼ AI-Helper

Tomato yellow leaf curl China virus is a species of the widespread geminiviruses. The infection of Nicotiana benthamiana by Tomato yellow leaf curl China virus (TYLCCNV) causes a reduction in photosynthetic activity, which is part of the viral symptoms. ${\beta}C1$ is a viral factor encoded by the betasatellite DNA ($DNA{\beta}$) accompanying TYLCCNV. It is a major viral pathogenicity factor of TYLCCNV. To elucidate the effect of ${\beta}C1$ on plants' photosynthesis, we measured the relative chlorophyll (Chl) content and Chl fluorescence in TY-LCCNV-infected and ${\beta}C1$ transgenic N. benthamiana plants. The results showed that Chl content is reduced in TYLCCNV A-infected, TYLCCNV A plus $DNA{\beta}$ (TYLCCNV A + ${\beta}$)-infected and ${\beta}C1$ transgenic plants. Further, changes in Chl fluorescence parameters, such as electron transport rate, $F_v/F_m$, NPQ, and qP, revealed that photosynthetic efficiency is compromised in the aforementioned N. benthamiana plants. The presense of ${\beta}C1$ aggravated the decrease of Chl content and photosynthetic efficiency during viral infection. Additionally, the real-time quantitative PCR analysis of oxygen evolving complex genes in photosystem II, such as PsbO, PsbP, PsbQ, and PsbR, showed a significant reduction of the relative expression of these genes at the late stage of TYLCCNV A + ${\beta}$ infection and at the vegetative stage of ${\beta}C1$ transgenic N. benthamiana plants. In summary, this study revealed the pathogenicity of TYLCCNV in photosynthesis and disclosed the effect of ${\beta}C1$ in exacerbating the damage in photosynthesis efficiency by TYLCCNV infection.

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표/그림 (6)

그림 Fig. 1. Tomato yellow leaf curl China virus (TYLCCNV) infection and βC1 transgene induce abnormal phenotype in Nicotiana benthamiana.
그림 Fig. 2. βC1 reduces the chlorophyll contents in Nicotiana benthamiana.
그림 Fig. 3. Chlorophyll fluorescence parameters of the Tomato yellow leaf curl China virus (TYLCCNV)-infected Nicotiana benthamiana plants. Chlorophyll fluorescence parameters in systemic (A) and inoculated (B) leaves of the untreated, mock-inoculated, TYLCCNV A and TYLCCNV A + β-infected N. benthamiana plants at 0-14-day post-inoculation (dpi).
그림 Fig. 4. Chlorophyll fluorescence images of the wild type and βC1 transgenic Nicotiana benthamiana leaves.
그림 Fig. 5. Tomato yellow leaf curl China virus (TYLCCNV) infection reduced the relative expression of major photosystem II (PSII) extrinsic genes.
그림 Fig. 6. βC1 transgene reduced the expression of oxygen evolving complex (OEC) genes at vegetative stage.

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제안 방법

The glyceradehyde 3-phosphate dehydrogenase (GAPDH) was used as the reference gene to calculate the relative expression of target genes. All RT-qPCR reactions were performed using three biological and nine technical repeats, and the data were subjected to statistical analysis. The specific primer pairs for RT-qPCR were designed by using PrimerQuest program (Integrated DNA Technologies, Coralville, IW, USA).
, 2016). In this study, we used Chl fluorescence imaging to probe the PSII activities during TYLCCNV infection. Obvious changes in fluorescence dynamics were observed in both TYLCCNV A– and TYLCCNV A + β–infected plants (Fig.
Following a dark adaptation of 30 min, plants were continuously kept in darkness during the period of fluorescence measurement. QY-max (equal to F_v/F_m), steadystate nonphotochemical quenching in light (NPQ-Lss) and steady-state fluorescence decline ratio in light (Rfd-Lss) were measured to evaluate the photosynthetic efficiency of the plants. False color was used to display the images.
Real-time quantitative PCR analysis was performed to determine the relative expression levels of the PSII genes, PsbO, PsbP, PsbQ, and PsbR, in untreated, mock-inoculated, TYLCCNV A and TYLCCNV A + β–infected Nicotiana benthamiana plants at 5- and 14-day post-inoculation (dpi).
The transcript levels of OEC genes were analyzed in the systemic leaves of untreated, mock-inoculated, TYLCCNV A– and TYLCCNV A+β–infected N. benthamiana plants using GAPDH as an internal standard for relative quantification.
To investigate the possible changes in transcription of these genes which might be induced by TYLCCNV infection or viral factor βC1, RT-qPCR analysis was performed for the 5 and 14 dpi samples.

이론/모형

As the needle-like structure of the βC1 transgenic N. benthamiana plants does not fit the leaf clip of SPAD, Arnon’s method was applied to measure the Chl content of these tissues.

성능/효과

The results showed that βC1 transgenic plants contained significantly lower amounts of Chl a, Chl b, and total Chl comparing to the wild type (Fig. 2C).
The results showed that the transcription levels of PsbO, PsbP, PsbQ, and PsbR remained unchanged at 5 dpi. However, a significant downregulation of these genes was observed in TYLCCNV A + β–infected plants at 14 dpi comparing to the untreated controls or plants with other treatments (Fig.
These results showed that TYLCCNV A alone triggered only a mild reduction in Chl content without obvious changes in the color of leaves, and the presence of DNAβ exacerbated the reduction.

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