Regulation of Ethylene Biosynthesis in Phytochrome Mutants of the Arabidopsis Root

Ji Hye Park(박지혜),Soon Young Kim(김순영) 한국생명과학회 2012 생명과학회지 Vol.22 No.4

식물생장과 발달에 중요한 역할을 하는 phytochrome이 ethylene 생합성에 미치는 영향을 조사하기 위하여 여러 빛 조건에서 키운 phyA, phyB, phyAB에서 ethylene 생합성과 생합성에 관여하는 enzyme activity를 측정하였다. White light에서 키웠을 때 모든 mutant에서 ethylene 생합성이 감소되었다. 특히 double mutant에서는 wild type과 비교하여 37%가 감소하였다. Dark에서 키웠을 때에는 wild type만 감소하였고, mutant에서는 감소효과가 나타나지 않았다. Red light에서 키웠을 때 double mutant에서 급격한 감소가 일어났다. Far-red light 에서 키웠을 때는 phyB만 감소가 일어나지 않았다. Ethylene 생합성에 관여하는 enzyme인 ACO 활성 패턴과는 달리ACS 활성 패턴은 ethylene 생성 패턴과 유사하게 나타났다. 이 결과를 바탕으로 ethylene 생합성에는 phytochromeA와 B 모두 중요한 작용을 하며 특히 Pr 형태의 phytochrome이 ethylene 생성량을 조절한다는 것을 제시한다. 또한 phytochrome은 ethylene 생합성 단계에서 AdoMet가 ACC로 전환되는 단계에서 조절하는 것을 제시한다. In order to investigate the effect of phytochromes on the regulation of ethylene biosynthesis, we measured the ethylene production and the activities of enzymes involved in ethylene biosynthesis using phytochrome mutants such as phyA, phyB, and phyAB of Arabidopsis. The ethylene production was decreased in mutants grown in white light. In particular, double mutants showed a 37% decrease compared to the wild type in ethylene production. When Arabidopsis roots were grown in the dark, mutants did not show a decrease in ethylene production; however, production was significantly decreased in the double mutant grown in red light. Only phyB did not show the decrease in the ethylene production in far-red light. Unlike the ACO activities, the ACS activities of mutants showed the same pattern as the ethylene production under several light conditions. The results of ACS activities confirmed the expression of the ACS gene by RT-PCR analysis. The decrease of ethylene production in mutants was due to the lower activity of ACC synthase, which converts the S-adenosyl-L-methionine (AdoMet) to 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene. These results suggested that both phytochrome A and B play an important role in the regulation of ethylene biosynthesis in Arabidopsis roots in the conversion step of AdoMet to ACC, which is regulated by ACS.

Phytochromes A and B: Specificity of photoperception and structure/function analysis of bilin chromophores

Shinomura, Tomoko,Hanzawa, Hiroko,Furuya, Masaki Korean Society of Photoscience 2002 Journal of Photosciences Vol.9 No.2

Phytochrome A (phyA) and phytochrome B (phyB) perceive light and adapt to fluctuating circumstances by different manners in terms of effective wavelengths, required fluence and photoreversibility. Action spectra for induction of seed germination and inhibition of hypocotyl elongation using phytochrome mutants of Arabidopsis showed major difference. PhyA is the principal photoreceptor for the very low fluence responses and the far-red light-induced high irradiance responses, while phyB controls low fluence response in a red/far-red reversible mode. The structural requirement of their bilin chromophores for photosensory specificity of phyA and phyB was investigated by reconstituting holophytochromes through feeding various synthetic bilins to the following chromophore-deficient mutants: hy1, hyl/phyA and hyl/phyB mutants of Arabidopsis. We found that the vinyl side-chain of the D-ring in phytochromobilin interacts with phyA apoprotein. This interaction plays a direct role in mediating the specific photosensory function of phyA. The ethyl side-chain of the D-ring in phycocyanobilin fails to interact with phyA apoprotein, therefore, phyA specific photosensory function is not observed. In contrast, both phytochromobilin and phycocyanobilin interact with phyB apoprotein and induce phyB specific photosensory functions. Structural requirements of the apoproteins and the chromophores for the specific photoperception of phyA and phyB are discussed.

Phytochromes are Involved in the Regulation of Growth and the Gravitropic Response via Ethylene Production in Hypocotyl of Arabidopsis

Sang Seung Lee(이상승),Soon Young Kim(김순영) 한국생명과학회 2018 생명과학회지 Vol.28 No.1

피토크롬은 빛을 인지하여 식물의 생장과 발달에 영향을 미치고, 식물호르몬인 에틸렌은 식물의 줄기 뿌리의 생장을 조절한다. 본 연구는 phyA, phyB and phyAB와 같은 애기장대의 피토크롬 돌연변이체를 이용하여 다양한 빛 조건(암소, white light, red light, far red light)에서 하배축의 생장과 굴중성 반응을 측정하였다. 모든 빛 조건에서 돌연변이체 phyAB는 다른 돌연변이체와 wild type (WT)보다 생장과 굴중성 반응이 가장 촉진되었다. Red light (R)에서 phyB가 phyA보다 굴중성 반응이 촉진되었으나 far red light (FR)에서는 phyB가 phyA보다 굴중성반응이 억제되었다. 하배축의 생장도 굴중성 반응과 같은 양상으로 조절되었다. 피토크롬의 작용을 설명하기 위하여 에틸렌 생성과 in vitro ACS, ACO 활성을 측정하였다. White light에서 돌연변이체보다 WT에서 에틸렌 생성이 촉진되었다. 그러나 R에서 키운 phyA와 FR에서 키운 phyB에서 에틸렌 생성이 촉진되어 WT와 비슷한 생성량을 보였다. ACS 활성도 에틸렌 생성량의 양상과 일치하였다. 이 결과는 R에서는 phyB의 Pr 형태가, 그리고 FR에서는 phyA의 Pfr 형태가 에틸렌 생성을 조절하여 하배축의 생장과 굴중성 반응을 조절한다는 가능성을 제시한다. Light is essential to the growth and development of plants, and it is perceived by phytochromes, which are one of the photoreceptors that regulate physiological responses in plants. Ethylene regulates the dormancy, senescence, growth, and development of organs in plants. This research focused on the interaction of phytochromes and ethylene to control hypocotyl growth and gravitropism using phytochrome mutants of Arabidopsis, phyA, phyB, and phyAB, under three light conditions: red (R) light, farred (FR) light, and white light. The mutant phyAB exhibited the most stimulation of gravitropic response of all three phytochrome mutants and wild type (WT) in all three light conditions. Moreover, phyB in the R light condition showed more negative gravitropism than phyA. However, phyB in the FR light condition showed less curvature than phyA. The hypocotyl growth pattern was similar to the gravitropic response in several light conditions. To explain the mechanism of the regulation of gravitropic response and growth, we measured the ethylene production and activities of in vitro ACS and ACO. Ethylene production was reduced in all the mutants grown in white light in comparison to the WT. Ethylene production increased in the phyA grown in R light and phyB grown in FR light in comparison to the other mutants. The ACS activity coincided with the ethylene production in the phyA and the phyB grown in R light and FR light, respectively. These results suggest that the Pfr form of phyB in R light and the Pr form of phyA in FR light increased ethylene production via increasing ACS activity.

Root Gravitropic Response of Phytochrome Mutant ( phyAB) in Arabidopsis

Soon Hwa Woo(우순화),Seung-Eun Oh(오승은),Jong Sik Kim(김종식),Jack L. Mullen,Roger P. Hangarter,Soon Young Kim(김순영) 한국생명과학회 2008 생명과학회지 Vol.18 No.2

Arabidopsis의 피토크롬 2중 돌연변이형 (phyAB)은 야생형 (WT)과 비교하여 뿌리의 굴중성 반응이 지연되었다. 중력 자극을 받은 지 8시간 후에 돌연변이체의 굴중성 반응은 야생형의 48%를 나타내었다. 지연된 반응은 중력자극을 준 후 1.5 시간 뒤에 나타났다. 12시간 동안 야생형과 돌연변이형의 뿌리 절편에서 에틸렌 생성을 측정하였다. 돌연변이형의 에틸렌 생성은 12시간이 경과한 후에 야생형의 40% 정도로 감소되었다. 이러한 결과는 피토크롬이 에틸렌 생성과 연관되어 있음을 제시하고 있다. 일반적으로 에틸렌은 식물의 뿌리나 줄기를 억제한다. 본 연구에서는 에틸렌 전구체인 1-aminocycloprpane-1-carboxylic acid (ACC)를 처리하여 뿌리의 생장을 측정하였다. 야생형은 ACC 존재하에 뿌리 생장이 억제되었으나, 돌연변이형은 야생형만큼 억제를 나타내지 않았다. 이 결과를 확인하기 위하여 ACC 존재 하에 굴중성 반응을 측정한 결과, 야생형은 ACC가 없는 경우와 비교하여 37.4%의 억제를 나타냈으나, 돌연변이형은 ACC가 없는 경우와 비교하여 6.6%만을 억제하였다. 이 결과는 피토크롬이 에틸렌 생성을 통하여 뿌리 굴중성 반응을 조절한다는 것을 제시한다. Phytochrome double mutant (phyAB) showed the delayed root gravitropic response compared to the wild type (WT) in Arabidopsis. After 8 hr of gravistimulation, the gravitropic response of mutant showed 48% of the WT. The delayed response started at 1.5 hr after gravistimulation. And we measured the ethylene production in the root segments of WT and mutant for 12 hr. Ethylene production of mutant decreased about 40% of the WT at 12 hr. This result suggested that the phytochrome might be linked with ethylene production in some way. Generally, ethylene inhibits the growth of plant organs including roots. We measured the root growth rate in the presence of ACC (1-aminocyclopropane-1-carboxylic acid), a precursor of ethylene. And WT showed the inhibition of root growth with ACC, but mutant did not show the inhibition as WT did. To confirm the relationship between the ethylene and gravitropic response, we measured the gravitropic response with ACC. In the presence of 10-6 M ACC, WT showed the 37.4% inhibition compared to the control (no ACC), whereas mutant showed the only 6.6% inhibition of control (no ACC). This research suggested the relationship between phytochrome and gravitropic response through an ethylene production.

Photo-biotechnology as a tool to improve agronomic traits in crops

Gururani, Mayank Anand,Ganesan, Markkandan,Song, Pill-Soon Elsevier 2015 Biotechnology advances Vol.33 No.1

<P><B>Abstract</B></P> <P>Phytochromes are photosensory phosphoproteins with crucial roles in plant developmental responses to light. Functional studies of individual phytochromes have revealed their distinct roles in the plant's life cycle. Given the importance of phytochromes in key plant developmental processes, genetically manipulating phytochrome expression offers a promising approach to crop improvement. Photo-biotechnology refers to the transgenic expression of phytochrome transgenes or variants of such transgenes. Several studies have indicated that crop cultivars can be improved by modulating the expression of phytochrome genes. The improved traits include enhanced yield, improved grass quality, shade-tolerance, and stress resistance. In this review, we discuss the transgenic expression of phytochrome A and its hyperactive mutant (Ser599Ala-PhyA) in selected crops, such as <I>Zoysia japonica</I> (Japanese lawn grass), <I>Agrostis stolonifera</I> (creeping bentgrass), <I>Oryza sativa</I> (rice), <I>Solanum tuberosum</I> (potato), and <I>Ipomea batatas</I> (sweet potato). The transgenic expression of PhyA and its mutant in various plant species imparts biotechnologically useful traits. Here, we highlight recent advances in the field of photo-biotechnology and review the results of studies in which phytochromes or variants of phytochromes were transgenically expressed in various plant species. We conclude that photo-biotechnology offers an excellent platform for developing crops with improved properties.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Phytochromes are photoreceptors that are required for various developmental processes. </LI> <LI> Application of engineered phytochrome genes in various crops is discussed. </LI> <LI> Results with introduction of a mutant PHYA into turfgrasses and other crops are discussed. </LI> <LI> Potential areas for further improvement in photo-biotechnology are highlighted. </LI> </UL> </P>

Photochemical Characterization of Phytochrome Missense Mutants

SHIN, Ah-Young,KIM, Hwan-Sik,LEE, Si-Seok,KIM, Min-Gon,HAN, Yun-Jeong,SONG, Pill-Soon,KIM, Jeong-Il Japan Society for Bioscience, Biotechnology, and A 2011 Bioscience, biotechnology, and biochemistry Vol.75 No.12

<P>Phytochromes are photoreceptors that regulate many aspects of plant growth and development in response to red/far-red light signals from the environment. In this study, we analyzed chromophore ligation and photochromism of missense phytochrome mutants in the Per-Arnt-Sim (PAS)-related domain (PRD). Among the 14 mutants analyzed, the Gly768Asp mutant of <I>Avena</I> phytochrome A showed aberrant photochromism and dark reversion, suggesting that amino acid residues in the C-terminal domain affect the photochemical properties of the photosensory N-terminal domain.</P>