Emerging Roles of RNA-Binding Proteins in Plant Growth, Development, and Stress Responses

Lee, Kwanuk,Kang, Hunseung Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.3

Posttranscriptional regulation of RNA metabolism, including RNA processing, intron splicing, editing, RNA export, and decay, is increasingly regarded as an essential step for fine-tuning the regulation of gene expression in eukaryotes. RNA-binding proteins (RBPs) are central regulatory factors controlling posttranscriptional RNA metabolism during plant growth, development, and stress responses. Although functional roles of diverse RBPs in living organisms have been determined during the last decades, our understanding of the functional roles of RBPs in plants is lagging far behind our understanding of those in other organisms, including animals, bacteria, and viruses. However, recent functional analysis of multiple RBP family members involved in plant RNA metabolism and elucidation of the mechanistic roles of RBPs shed light on the cellular roles of diverse RBPs in growth, development, and stress responses of plants. In this review, we will discuss recent studies demonstrating the emerging roles of multiple RBP family members that play essential roles in RNA metabolism during plant growth, development, and stress responses.

저온 및 고온내성 고추 우량 계통 조기선발을 위한 발아 및 유묘 시험 결과

이관욱(Kwanuk Lee),조명철(Myeong Cheol Cho),양은영(Eun Young Yang),정효봉(Hyo Bong Jeong),쉐르조드(Rajametov Sherzod),남춘우(Choon Woo Nam),김정호(Jeong Ho Kim) 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.10

고추는 노지와 하우스 내에서 연중 재배되고 있으며, 생물학적와 무생물학적 스트레스를 포함한 많은 환경스트레스에 노출되어 있다. 최근 기후 변화에 따른 잦아진 저온 및 고온 이상기상은 고추의 생육과 발달에 큰 영향을 미치며, 생리장해, 낙화, 낙과, 착과불량, 수확량 감소로 이어져 농가 소득 증감에 영향을 미치고 있다. 따라서, 저온 및 고온 적응성이 우수한 고추 우량계통을 조기 선발하여 농가에 보급하는 것이 중요하며, 본 연구에서는 저온 및 고온내성 고추 우량 계통 조기선발을 위한 발아기 및 유묘기 실험을 진행하였다. 고추 ‘녹광’과 ‘청양’ 품종의 종자를 이용하여 발아기 단계에서 저온 (15°C), 정상온도(25°C), 그리고 고온(35°C)을 처리하였으며, 4일째 발아세와 7일째 발아율을 측정하였다. 저온(15°C)에서 측정한 고추 ‘녹광’과 ‘청양’의 종자발아세는 0%와 35.3%, 발아율은 0%와 73.9%로 ‘청양’의 발아세와 발아율이 우수하였다. 정상온도(25°C)에서 녹광’과 ‘청양’ 종자발아세는 59.3%와 26.8%, 발아율은 100%와 73.2%로 ‘녹광’ 종자발아세와 발아율이 ‘청양’보다 높은 특성을 보였다. 고온(35°C)에서 고추 ‘녹광’과 ‘청양’의 발아세는 21.6%와 28.8%, 발아율은 75.8%와 29.9%로 종자발아세는 비슷하였지만, ‘녹광’ 종자발아율이 더 높은 특성을 보였다. 10 ‒ 12일 생장한 유묘기 단계 식물에 저온(2°C), 정상온도(25°C), 그리고 고온(42°C)을 16시간 처리하여, 생존률(%), 엽록소 함량, ROS 함량을 조사하였다. 그 결과 저온과 정상온도 처리 유묘의 뚜렷한 차이가 없었고, 고온처리 후 ‘녹광’ 유묘의 생존률이 ‘청양’ 보다 79% 정도 높았다. 또한, 고온 처리 한 유묘의 엽록소(chl a+b) 함량은 ‘녹광’에서 75%, ‘청양’에서 52% 유지되었고, ‘녹광’이 ‘청양’보다 23% 정도 높은 엽록소 함량을 보였다. 고온 처리한 식물체 내 ROS 함량 변화를 DAB staining을 통해 수행한 결과, 생존률이 높았던 ‘녹광’ 유묘에서 ‘청양’보다 더 낮은 ROS를 검출하였다. 이런 결과들은 저온 및 고온에서 고추 종자발아와 유묘 적응에 필요한 온도 범위가 다양하다는 것을 의미하고, 더 많은 고추 계통들의 발아기 및 유묘기 검정을 통해 저온 및 고온내성 우량계통 선발 및 육종자원으로 활용될 것으로 기대한다.

저온 및 고온내성 토마토, 고추 우량 계통 조기선발을 위한 발아 시험 결과

이관욱(Kwanuk Lee),조명철(Myeong Cheoul Cho),양은영(Young Yang),정효봉(Hyo Bong Jeong),쉐르조드(Rajametov Sherzod),남춘우(Choon Woo Nam),김정호(Jeong Ho Kim) 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.5

토마토, 고추는 노지와 시설 내에서 재배되고 있으며, 생물학적, 무생물학적 스트레스를 포함한 다양한 환경스트레스에 노출되어 있다. 최근 기후 변화에 따른 잦아진 저온 및 고온 이상기상은 토마토와 고추의 생육과 발달에 영향을 미치며, 생리장해, 낙화, 낙과, 착과불량, 열과 발생으로 수확량 감소 및 과실 품질저하로 이어져 농가 피해가 급증가하고 있다. 저온 및 고온 적응성이 우수한 토마토, 고추 우량계통을 조기 선발하여 농가에 보급하는 것이 중요하며, 본 연구에서는 저온 및 고온내성 토마토, 고추 우량 계통 조기선발을 위한 발아 실험을 수행하였다. 토마토는 시판종인 ‘대프니스’와 ‘미니찰’을, 고추는 ‘녹광’과 ‘청양’을 이용하여 발아기 단계에서 저온 (15°C), 정상온도(25°C), 그리고 고온(35°C)을 처리하였으며, 4일째 발아세와 7일째 발아율을 측정하였다. 저온(15°C)에서 측정한 토마토 ‘대프니스’와 ‘미니찰’의 종자발아세는 0%와 7.2%, 발아율은 31.4%와 91.5%로 ‘미니찰’ 품종의 종자발아세와 발아율이 높은 특성을 보였다. 고추 ‘녹광’과 ‘청양’의 종자발아세는 0%와 35.3%, 발아율은 0%와 73.9%로 ‘청양’의 발아세와 발아율이 우수하였다. 정상온도(25°C)에서 측정한 ‘대프니스’와 ‘미니찰’ 종자발아세는 96.1%와 93.5%, 발아율은 98.7%와 100%로 비슷하였다. ‘녹광’과 ‘청양’ 종자발아세는 59.3%와 26.8%, 발아율은 100%와 73.2%로 ‘녹광’ 종자발아세와 발아율이 ‘청양’보다 우수한 특성을 보였다. 한편, 고온(35°C)에서 토마토 ‘대프니스’와 ‘미니찰’ 종자발아세는 0%와 2.6%, 발아율은 0%와 3.9%로 비슷하였다. 고추 ‘녹광’과 ‘청양’의 발아세는 21.6%와 28.8%, 발아율은 75.8%와 29.9%로 종자발아세는 비슷하였지만, ‘녹광’ 종자발아율이 더 우수한 특성을 보였다. 이 결과는 토마토와 고추 품종 간 종자 발아에 필요한 적응온도 범위가 많이 다름을 의미한다. 이러한 기초 연구를 기반으로 추후 다양한 토마토, 고추 품종과 계통을 이용한 발아 실험을 수행하고, 계통들의 포장에서 원예적 특성 평가를 통해 저온 및 고온내성 토마토, 고추 우량계통 선발에 활용할 수 있을 것으로 기대한다.

A chloroplast-targeted cabbage DEAD-box RNA helicase BrRH22 confers abiotic stress tolerance to transgenic <i>Arabidopsis</i> plants by affecting translation of chloroplast transcripts

Nawaz, Ghazala,Lee, Kwanuk,Park, Su Jung,Kim, Yeon-Ok,Kang, Hunseung Elsevier 2018 Vol. No.

<P><B>Abstract</B></P> <P>Although the roles of many DEAD-box RNA helicases (RHs) have been determined in the nucleus as well as in cytoplasm during stress responses, the importance of chloroplast-targeted DEAD-box RHs in stress response remains largely unknown. In this study, we determined the function of BrRH22, a chloroplast-targeted DEAD-box RH in cabbage (<I>Brassica rapa</I>), in abiotic stress responses. The expression of <I>BrRH22</I> was markedly increased by drought, heat, salt, or cold stress and by ABA treatment, but was largely decreased by UV stress. Expression of BrRH22 in <I>Arabidopsis</I> enhanced germination and plantlet growth under high salinity or drought stress. BrRH22-expressing plants displayed a higher cotyledon greening and better plantlet growth upon ABA treatment due to decreases in the levels of <I>ABI3, ABI4,</I> and <I>ABI5</I>. Further, BrRH22 affected translation of several chloroplast transcripts under stress. Notably, BrRH22 had RNA chaperone function. These results altogether suggest that chloroplast-transported BrRH22 contributes positively to the response of transgenic <I>Arabidopsis</I> to abiotic stress by affecting translation of chloroplast genes via its RNA chaperone activity.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Expression of cabbage RNA helicase BrRH22 is differentially affected by abiotic stresses. </LI> <LI> BrRH22 is localized to chloroplasts and possesses RNA chaperone activity. </LI> <LI> BrRH22 plays a positive role in seed germination and seedling growth under stress conditions. </LI> <LI> BrRH22 enhances seedling growth and cotyledon greening by decreasing <I>ABI3, ABI4,</I> and <I>ABI5.</I> </LI> </UL> </P>

Impact of Agrobacterium-infiltration and transient overexpression of BroMYB28 on glucoraphanin biosynthesis in broccoli leaves

Kim Young-Cheon,차아라,Hussain Muhammad,Lee Kwanuk,이상협 한국식물생명공학회 2020 Plant biotechnology reports Vol.14 No.3

To date studies revealed that MYB28 intensively involved biosynthesis of GSLs, there is no direct evidence of MYB28 in glucoraphanin (GR) biosynthesis of broccoli. Furthermore, Agrobacterium (Agro)-infiltration also induced GSLs in Arabidopsis. However, there is no information regarding Agro-dependent GR accumulation in broccoli. We revealed that over-expression of BroMYB28 could accumulate GR, and Agro-infiltration is also enough to induce the accumulation of GR in broccoli. From our results, it is suggested that they used different regulatory mechanisms; BroMYB28 over-expression induced both Bro- MYB28 and BroMYB29 (Bol008849-like) transcripts; Agro-infiltration induced only BroMYB29 (Bol008849-like) transcripts.

<i>quatre-quart1</i> is an indispensable U12 intron-containing gene that plays a crucial role in Arabidopsis development

Kwak, Kyung Jin,Kim, Bo Mi,Lee, Kwanuk,Kang, Hunseung Oxford University Press 2017 Journal of experimental botany Vol.68 No.11

<▼1><P><I>quatre-quart1</I> is an indispensable U12 intron-containing gene, and correct splicing of the U12 intron in <I>quatre-quart1</I> is crucial for the normal development of <I>Arabidopsis thaliana</I>.</P></▼1><▼2><P><B>Abstract</B></P><P>Despite increasing understanding of the importance of the splicing of U12-type introns in plant development, the key question of which U12 intron-containing genes are essential for plant development has not yet been explored. Here, we assessed the functional role of the <I>quatre-quart1</I> (<I>QQT1</I>) gene, one of the ~230 U12 intron-containing genes in <I>Arabidopsis thaliana</I>. Expression of <I>QQT1</I> in the U11/U12-31K small nuclear ribonucleoprotein mutant (<I>31k</I>) rescued the developmental-defect phenotypes of the <I>31k</I> mutant, whereas the miRNA-mediated <I>qqt1</I> knockdown mutants displayed severe defects in growth and development, including severely arrested stem growth, small size, and the formation of serrated leaves. The structures of the shoot apical meristems in the <I>qqt1</I> mutants were abnormal and disordered. Identification of QQT1-interacting proteins via a yeast two-hybrid screening and a firefly luciferase complementation-imaging assay revealed that a variety of proteins, including many chloroplast-targeted proteins, interacted with QQT1. Importantly, the levels of chloroplast-targeted proteins in the chloroplast were reduced, and the chloroplast structure was abnormal in the <I>qqt1</I> mutant. Collectively, these results provide clear evidence that <I>QQT1</I> is an indispensable U12 intron-containing gene whose correct splicing is crucial for the normal development of Arabidopsis.</P></▼2>

A La-Related Protein LaRP6a Delays Flowering of Arabidopsis thaliana by Upregulating FLC Transcript Levels

Park Su Jung,Hwa Jung Lee,Kwanuk Lee,강훈승 한국식물학회 2020 Journal of Plant Biology Vol.63 No.5

La-related proteins (LaRPs) are RNA-binding proteins (RBPs) that contain a La-type helix-turn-helix (HTH) domain and are found in all eukaryotes, including plants, animals, and yeasts. Although the functions of a few LaRPs have been determined during plant growth and development, the roles of most LaRPs in plants remain unknown. Here, the function of an Arabidopsis thaliana LaRP6a (At5g46250) harboring a La-type HTH domain and an RNA-recognition motif was determined in the growth and development of Arabidopsis. Confocal analysis of subcellular localization of the LaRP6a–GFP fusion protein revealed that LaRP6a protein is localized to both the nucleus and plasma membrane. Overexpression of LaRP6a resulted in delayed flowering and increased plant height and seed yield. The transcript level of the floral repressor FLC was markedly increased in LaRP6a-overexpressing transgenic plants. LaRP6a protein bound to the 3ʹUTR of the FLC transcript and possessed RNA chaperone activity. Collectively, these results suggest that LaRP6a plays a role in the control of flowering time by regulating FLC transcript levels.

Rice DEAD-box RNA helicase OsRH53 has negative impact on Arabidopsis response to abiotic stresses

Nawaz, Ghazala,Sai, Than Zaw Tun,Lee, Kwanuk,Kim, Yeon-Ok,Kang, Hunseung Springer-Verlag 2018 Plant growth regulation Vol.85 No.1

Comparison of Growth and Quantity by Varieties of Red Tomatoes according to Grafting or Non-grafting in the Four-season House

Myeong Cheoul Cho,Rajametov Sherzod,Hyo Bong Jeong,Eun Young Yang,Kwanuk Lee,Chun Woo Nam,Doo Jong Ha,Dan Hye Kim,Hyoung Je Yoo 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.10

The purpose of this study was to investigate effect of the grafting. We used “Spider” (Takii seed, Japan) rootstock and ‘Dokia’, ‘TY Red 250’ and ‘Pilabi’ varieties having red color at the four season house in South Korea. All tomato cultivars seedlings were transplanted on 4 March of 2021 to greenhouse, where day and night temperatures were maintained at 18/32°C, and relatively humidity was within D/N 40 ‒ 90%. Tomato seedlings grafted (GF) and non-grafted (control, CT) having the first truss were planted on substrate perlite (mixture of perlite no.1 and no.3), fertilization - EC 3.0ds/m, pH 6.0, 6 to12 times per day, cooling system-Forced ventilation (open above 25°C), cooling through fog system (on above 28°C). Drip irrigation systems are used (200 ‒ 250 mL per plant in each time). Plant height (PH), leaf length and width (LL and LW), days to flowering (DF), fruit weight (FW) and yield (FY) parameters were measured. Effect of grafting on vegetative and reproductive parameters of tomato was differed among cultivars. There were identified the significant increasing of the PH in GF plants of cv. ‘Dokia’ than CT for 150 days after transplanting (DAT), while cv. ‘TY Red 250’ showed opposite trend, it is decreased in GF plants until 90 DAT then showed no difference. Only in plants of cv. ‘Pilabi’ no found difference between GF and CT during all growth stages. The grafting contributed to delay the blossom significantly in 2nd truss among all tomato cultivars than CT, but in subsequent growth stages from 5th to 15th truss no identified significant difference DF, except cv. ‘TY Red 250’ in 10th truss. The leaf parameters (LL and LW) showed decreasing with aging of plants among all cultivars on 150 DAT and no observed significant difference during all growth period, except cv. ‘Dokia’ within 2nd and 15th truss. Also, with aging of plants in fruits from 10th truss were identified significantly decreasing of FW among all cultivars regardless of the treatments, but there had been persist the difference in values among cultivars and treatments. The significant high yield (the sum from 1st to 10th truss) were pickup from GF plants of cv. “TY Red 250” than in CT, whereas in cv. ‘Dokia’ and ‘Pilabi’ no found difference in yield between GF and CT plants.

Comparison of Growth and Quantity by Varieties of Pink Tomatoes according to Grafting or Non-grafting in the Four-season House

Myeong Cheoul Cho,Rajametov Sherzod,Hyo Bong Jeong,Eun Young Yang,Kwanuk Lee,Chun Woo Nam,Doo Jong Ha,Dan Hye Kim,Hyoung Je Yoo 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.10

The objective of this study was to evaluate effect of the grafting. We used ‘Spider’ (Takii seed, Japan) rootstock and big size (Pink type) tomato cultivars ‘Pink Star’, ‘K-Star’ and ‘Dotaerang’ at the four season house in South Korea. Bacterial wilt (caused by Ralstonia solanacearum) and Fusarium wilt (Fusarium oxysporum) diseases tolerant rootstock “Spider” was used. All tomato cultivars seedlings were transplanted with distance 25 cm of plants, in 4 March of 2021 to greenhouse, where day and night temperatures were maintained at 18/32°C, and relatively humidity was within D/N 40 ‒ 90%. Tomato seedlings grafted (GF) and non-grafted (control, CT) were planted on substrate perlite (mixture of perlite no.1 and no.3), fertilization - EC 3.0ds/m, pH 6.0, 6 to12 times per day, cooling system- Forced ventilation (open above 25°C), cooling through fog system (on above 28°C). Drip irrigation systems are used (200 ‒ 250 mL per plant in each time). Plant height (PH), leaf length and width (LL and LW), days to flowering (DF), fruit weight (FW) and yield (FY) parameters were measured. Effect of rootstock on vegetative and reproductive parameters of tomato was differed among cultivars. So, grafted plant significantly decreased growth rate among plants of cv. ‘K-Star’ for all growth period, and the same pattern were observed in cv. ‘Dotaerang’ and ‘Pink Star’ but it persists until 60 DAT (days after transplanting), respectively. Evaluation of influence of grafting on days to flowering among cultivars showed that there no identified significant difference starting of the flowering between GF and CT treatments for all growth period. Almost the same pattern was observed in study of LL and LF among all cultivars, where with aging of plants were identified decreasing of the LL and LF parameters regardless of cultivars and treatments. As mentioned above, with aging of plants were identified significantly decreasing of FW among all cultivars regardless of the treatments, but there was persist the difference in values among cultivars and treatments. And, among all cultivars no found significant difference in FW between GF and CT plants. The significant high yield per plant among cultivars were harvested from GF plants of cv. ‘Pin Star’, however there no found significant difference between GF and CT plants inside of each cultivar. It means GF no significantly affect on FW and FY parameters among studied pink type tomato cultivars.