[논문]Oryzalin이 애기장대 뿌리 생장과 굴중성 반응에 미치는 작용

고진규; 박선일; 김순영

doi:10.5352/jls.2021.31.2.109

Oryzalin이 애기장대 뿌리 생장과 굴중성 반응에 미치는 작용
The Effect of Oryzalin on Growth and Gravitropism in Arabidopsis Roots 원문보기

생명과학회지 = Journal of life science, v.31 no.2, 2021년, pp.109 - 114

고진규 (안동대학교 생명공학부 생명과학전공) , 박선일 (안동대학교 생명공학부 생명과학전공) , 김순영 (안동대학교 생명공학부 생명과학전공)

초록
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Oryzalin은 미세소관의 배열을 방해하는 dinitroaniline계 제초제이다. 미세소관과 미세섬유는 식물 세포 생장에 관여하는 세포벽을 구성하는 골격 성분이다. 또한 미세소관은 평형석의 침전에도 관여하는데, 이는 뿌리 끝에 있는 columella 세포에서 중력 인지를 조절한다. 본 연구는 애기장대 뿌리에서 에틸렌 생성을 통하여 oryzalin이 뿌리 생장과 굴중성 반응에 미치는 영향을 조사하였다. 10-4 M oryzalin을 뿌리에 처리하면 뿌리 생장과 굴중성 반응이 완전히 억제되었다. 뿌리 생장과 굴중성 반응을 조절하는 oryzalin의 작용을 알아보기 위해 애기장대 뿌리 절편에서 ethylene 생합성을 측정하였다. Oryzalin을 처리하면 ACC oxidase와 ACC synthase을 활성을 촉진하여 에틸렌 생성이 촉진되고, 증가된 ethylene은 미세소관의 배열을 변화시켜 뿌리 생장과 굴중성 반응을 억제한다.

Abstract ▼ AI-Helper

Oryzalin is a dinitroaniline herbicide that has been known to disrupt microtubules. Microtubules and microfilaments are components of cytoskeletons that are implicated in plant cell growth, which requires the synthesis of cellulose when cell walls elongate. In addition, microtubules are also involved in the sedimentation of statoliths, which regulate the perception of gravity in the columella cells of root tips. In this study, we investigated the effect of oryzalin on the growth and gravitropic response of Arabidopsis roots. The role of ethylene in oryzalin's effect was also examined using these roots. Treatment of oryzalin at a concentration of 10-4 M completely inhibited the roots' growth and gravitropic response. At a concentration of 10-6 M oryzalin, root growth was inhibited by 47% at 8 hr when compared to control. Gravitropic response was inhibited by about 38% compared to control in roots treated with 10-6 M oryzalin for 4 hr. To understand the role of oryzalin in the regulation of root growth and gravitropic response, we measured ethylene production in root segments treated with oryzalin. It was found that the addition of oryzalin stimulated ethylene production through the activation of ACC oxidase and ACC synthase genes, which are key components in the synthesis of ethylene. From these findings, it can be inferred that oryzalin inhibits the growth and gravitropic response of Arabidopsis roots by stimulating ethylene production. The increased ethylene alters the arrangement of the microtubules, which eventually interferes with the growth of the cell wall.

주제어

표/그림 (6)

그림 Fig. 1. Effect of oryzalin on root growth in Arabidopsis seed-lings for 8 hr. After a vernalization for 1 day, seeds were grown for 6 days under diurnal rhythm of day and night cycle with 16 hr and 8 hr, respectively. These seedlings were transferred the agar plate containing several concentrations of oryzalin, and placed in vertical position. The growth was measured for 8 hr using a camera as described in Material and Methods. Symbols are mean values ± SE from 20 independent experiments.
표 Table 1. Gene-specific primers used for RT-PCR experiments. ACO2 and ACS are primers for the ACC oxidase and ACC synthase gene from Arabidopsis thaliana root, respectively
그림 Fig. 2. Effect of oryzalin on root gravitropic response in Arabid-opsis seedlings for 8 hr. After a vernalization for 1 day, seeds were grown for 6 days under diurnal rhythm diur-nal rhythm of day and night cycle with 16 hr and 8 hr, respectively. These seedlings were transferred the agar plate containing several concentrations of oryzalin, and placed in horizontal position to give a gravity. The gravitropic curvature was measured for 8 hr using a cam-era as described in Material and Methods. Symbols are mean values ± SE from 20 independent experiments.
그림 Fig. 3. Effect of oryzalin on the ethylene production in root segment for 8 hr. Root segments (10 mm) were in-cubated in solution containing 10^-8 M, 10^-6 M and 10^-4 M oryzalin. At every 2 hr, 1 ml of gas sample was withdrawn from the vials for measuring the production of ethylene. Symbols are mean values ± SE from 15 in-dependent experiments.
그림 Fig. 4. Effect of oryzalin on in vitro ACS and in vitro ACO activities. To measure in vitro ACS activity, one hun-dred of roots was incubated for 4 hr in solution contain-ing 10^-8 M, 10^-6 M and 10^-4 M oryzalin. Phosphate buffer (250 mM) was added to ground root segments in liquid nitrogen, and then ethylene production was measured as described in Materials and Methods. To measure in vitro ACO activity, one hundred of roots were in-cubated with oryzalin for 4 hr. These root segments were ground in liquid nitrogen, and added to an ex-traction buffer. After centrifugation, the supernatant was transferred to a new vial containing incubation buffer. Ethylene production was measured as described in Material and Methods to detect ACO activity. Symbols are mean values ± SE from 10 independent experiments.
그림 Fig. 5. Effect of oryzalin on gene expression of ACO and ACS. Root segments were incubated for 4 hr in solution con-taining 10^-8 M, 10^-6 M and 10^-4 M oryzalin. Total RNAs were extracted and used for RT-PCR as described in Material and Methods. Gel image show result with con-sistent patterns from 3 independent experiments.

AI 본문요약
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문제 정의

In this study, we found that the treatment of oryzalin interrupted the root growth and gravitropism and increased ethylene synthesis. So, it is considered that the increased ethylene induced by oryzalin affected the assembly and re-orientation of microtubules, which consequently inhibited the normal growth and gravitropic response of roots.

대상 데이터

The sterilized seeds of Landsberg erecta (Ler), Arabidopsis thaliana were planted on the agar medium with half-strength of MS salts, 1% sucrose and 1 mM MES (pH 5.8). The seeds were incubated in vertical position at 4℃ for 1 day and then were incubated for another 6 days at 22℃.

데이터처리

To test for significance at p values of <0.05, the data mean values were calculated according to one-way ANOVA and Tukey test.

성능/효과

In conclusion, the inhibition of growth and reduction of gravitropic response in Arabidopsis root by the treatment of oryzalin was mediated via the stimulation of ethylene biosynthesis, which was attributed to the increased ACO and ACS activities. The results found in this study are well consistent with the previous findings that ethylene changes the arrangement of microtubules, leading to inhibition of root growth and reduction of gravitropic response.
In conclusion, the inhibition of growth and reduction of gravitropic response in Arabidopsis root by the treatment of oryzalin was mediated via the stimulation of ethylene biosynthesis, which was attributed to the increased ACO and ACS activities. The results found in this study are well consistent with the previous findings that ethylene changes the arrangement of microtubules, leading to inhibition of root growth and reduction of gravitropic response.
To distinguish whether the increased ACO and ACS activities was due to the increase of transcript levels of the respective genes, RT-PCR was carried out with total RNAs extracted from roots treated with or without oryzalin (Fig. 5).

참고문헌 (18)

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