CURLY LEAF Research Articles

Sequencing CURLY LEAF-associated RNAs in Arabidopsis revealed prevalent intergenic RNAs from the nuclear mitochondrial sequence

Polycomb Group (PcG) proteins play key roles in development by repressing thousands of targets through histone modifications. However, how PcG is recruited to specific targets is poorly understood. In Arabidopsis, certain noncoding RNAs are necessary for recruiting the PcG protein CURLY LEAF (CLF) to its target sites. However, RNAs associated with CLF have not been analyzed on a genomic scale, thus it is unknown whether long noncoding RNA (lncRNA)-mediated PcG recruitment is a widespread mechanism. Here, we systematically searched for CLF-associated RNAs by RNA immunoprecipitation followed by deep sequencing. We identified 1299 genic and 138 intergenic regions that produced CLF-associated mRNAs and putative lncRNAs, respectively. The genes producing CLF-associated RNAs are depleted in PcG targets, carry active chromatin marks, and are highly expressed, suggesting that CLF may have a non-specific or promiscuous RNA-binding affinity, similar to animal PcG proteins. Notably, a significant portion of the CLF-associated lncRNAs is derived from the nuclear mitochondrial sequence, which is extensively marked by H3K27me3. These findings indicate that while CLF-RNA interactions are widespread, they may not always correlate with PcG target sites, highlighting the complexity of PcG recruitment mechanisms in Arabidopsis.

Isolation and identification of Cladosporium tenuissimum causing leaf blight on garden pea (Pisum sativum L.) in Telangana from southern India

Garden pea (Pisum sativum L.), is an important legume grown worldwide as a source of proteins and carbohydrates. Since pea crop is cultivated globally in cool temperate climatic conditions, increasing temperatures accompanied by high humidity favors fungal infections. A fungal infection causing leaf blight symptoms was frequently observed on the leaves of pea plants grown in greenhouse and field conditions. The blight symptoms showed a water-soaked and wilting appearance with curly leaf edges accompanied by greyish mycelial growth and chlorotic lesions. The disease incidence in field conditions ranged between 5 and 10 %, whereas in the controlled greenhouse conditions, it ranged from 40 to 45%. Two isolates of the fungus, Gp03 and Gp04 were isolated from Arkel pea leaves and purified from a single conidium each. According to Koch's postulates, the pathogenicity tests with these isolates using detached leaf assay and whole plant leaf assays confirmed the pathogen. Morphological and molecular analysis of nucleotide sequences of the D1-D2 region, translation elongation factor-1α (TEF) and actin (ACT) genes confirmed these isolates as members of the genus, Cladosporium. The phylogenetic relatedness of these isolates with other members of the Cladosporium genus revealed them as fungal strains of Cladosporium tenuissimum. Further, these isolates were also confirmed as C. tenuissimum by Microbial Type Culture Collection and Gene Bank (MTCC), Chandigarh in India, and provided the accession nos. MTCC 13581 and MTCC 13582 to the C. tenuissimum strains, Gp03 and Gp04 respectively. This is the first report of Cladosporium tenuissimum infecting garden pea in Telangana, India.

Brassica rapa CURLY LEAF is a major H3K27 methyltransferase regulating flowering time

Main conclusionIn Brassica rapa, the epigenetic modifier BraA.CLF orchestrates flowering by modulating H3K27me3 levels at the floral integrator genes FT, SOC1, and SEP3, thereby influencing their expression.CURLY LEAF (CLF) is the catalytic subunit of the plant Polycomb Repressive Complex 2 that mediates the trimethylation of histone H3 lysine 27 (H3K27me3), an epigenetic modification that leads to gene silencing. While the function of CURLY LEAF (CLF) has been extensively studied in Arabidopsis thaliana, its role in Brassica crops is barely known. In this study, we focused on the Brassica rapa homolog of CLF and found that the loss-of-function mutant braA.clf-1 exhibits an accelerated flowering together with pleiotropic phenotypic alterations compared to wild-type plants. In addition, we carried out transcriptomic and H3K27me3 genome-wide analyses to identify the genes regulated by BraA.CLF. Interestingly, we observed that several floral regulatory genes, including the B. rapa homologs of FT, SOC1 and SEP3, show reduced H3K27me3 levels and increased transcript levels compared to wild-type plants, suggesting that they are direct targets of BraA.CLF and key players in regulating flowering time in this crop. In addition, the results obtained will enhance our understanding of the epigenetic mechanisms regulating key developmental traits and will aid to increase crop yield by engineering new Brassica varieties with different flowering time requirements.

Viral complexity of tomatoes yellow mosaic and leaf curl diseases in lowland and highland areas

AbstractYellow mosaic and leaf curl diseases caused by whitefly‐transmitted begomo virus genera are important problems in tomato (Solanum lycopersicon L.) production in Indonesia, particularly on Java Island. The aims of this survey are to assess the percentages of disease incidence (DI) and disease severity index (DSI), evaluate the spread and complexity of begomovirus and crinivirus infections responsible for yellow mosaic and leaf curl diseases in tomato plants, and identify the species of whitefly insect vectors and their cryptic biotypes in tomato production centers located on Java Island, Indonesia. A total of 43 locations in East, Central, and West Java were surveyed during the dry season from July 2022 to September 2022. Disease incidence and severity were determined by observing 200 samples of tomato plants from five points at each location, with 40 plants per point of land. A total of 116 whitefly samples in microtubes and 215 symptomatic leaf samples were collected. The genus begomovirus and crinivirus infecting tomato were identified by PCR and RT‐PCR using genus‐specific primers. Similarly, whitefly species and their cryptic biotypes were identified by PCR using MCoI gene‐specific primers. Results showed that DI and DSI remarkably differed between the lowlands (66%–89%) and highlands (33%–62%) in the provinces of Central Java and West Java. Furthermore, PCR analysis of 215 infected tomato showing the symptoms of yellow mosaic and curly leaf revealed that three begomoviruses, one crinivirus and their mixed combination both begomoviruses, one crinivirus has been detected in all the samples. Similarly, the whitefly insect vector Bemisia tabaci biotype Q was identified in all areas, whereas Trialeurodes vaporariorum was found only in the East Java and Java highlands, respectively.

First Report of Globisporangium (Pythium) mastophorum causing Damping-off / Root Rot on Parsley in Slovenia.

Pythium-like species cause damping-off symptoms of various hosts, including umbelliferous crops. In April 2023, parsley plantlets (Petroselinum crispum), showing stunted growth, yellowing, decayed roots and damping-off, were obtained from a nursery in central Slovenia, where parsley was grown in polystyrene trays in a greenhouse. Nearly 30% of plants were symptomatic. Sampled roots of ten plants contained ornamented oogonia (avg. 33.3 ± 1.4 µm in diam) with conical projections (5.2 ± 0.5 µm long) (Figure S1 A, B) in microscopically analyzed squash mounts. The pathogen was isolated from root pieces treated for surface disinfection with 0.5% sodium hypochlorite for 30 s, and washed with sterile water. Four 1-2 mm root pieces were taken from each of 10 plants, plated on the selective medium P5ARP, and incubated at 21 °C. Mycelia emerging from root pieces were transferred to carrot piece agar (CPA). Twenty-two equally looking oomycetous colonies were obtained; all sampled plants were infested. Oogonia formed by all colonies were similar to those observed on decayed roots and suggested that Globisporangium (Pythium) mastophorum is the causal disease agent. Analyses of partial β-tubulin (Kroon et al. 2004) and mitochondrial cytochrome c oxidase I (COI) gene sequences (Robideau et al. 2011) confirmed the identification. Obtained COI (Genbank accession number OR725417) sequence was 100% identical to that from G. mastophorum strain CBS 375.72 (EU350523), whereas the β-tubulin sequence (OR725416) corresponded to 99.6 % pairwise identity (KJ595502). Further, pathogenicity of an obtained isolate was tested on 4 wk-old curly leaf (cv. Petra F1) parsley. Half of a 7 d-old CPA culture, consisting of mycelium and oogonia, was finely cut and mixed with ca 50 ml of nonsterile commercial substrate (Potgrond H, AGRO-FertiCrop) in each of six 400 ml pots. Pots were filled with ca 300 ml additional substrate, into which 5 parsley seedlings were planted. Control plants were treated equally but with sterile CPA. Plantlets were watered with sterile tap water and held at ambient light conditions and temperature (night 18 °C - day 23 °C). After 14 d, inoculated plants started wilting and yellowing and showed stunted growth. After 21 d, roots were severely decayed and the seedlings damped-off (Figure S1 C). Four pieces each from 10 decayed roots were plated. Thirty-one pieces revealed pythium-like colonies. Obtained isolates were morphologically identical to the strain used for inoculation and identified as G. mastophorum. Control plants developed no foliar or root symptoms and no pythium-like species was obtained. Agricultural advisors observed occurrence of parsley damping-off also in other nurseries in Slovenia what may lead to spreading the pathogen to parsley in production fields and private gardens. The case emphasizes the need for implementing phytosanitary measures in order to eliminate primary inoculum. Reports from field-infected plants showed that G. mastophorum is a pathogen of parsley in Australia (Petkowski et al. 2013) and the USA (Tsuchida et al. 2018), and celery in the Czech Republic (Šafránková and Holková 2017). Others isolated G. mastophorum from parsley in The Netherlands (online database of the Westerdijk Fungal Biodiversity Institute, strain CBS 243.86). However, the here described case is, to the best of our knowledge, one of the rare documentations of damping-off due to G. mastophorum in Europe (Šafránková and Holková 2017) and the first in Slovenia. Funding: The work was funded by the Ministry of Agriculture, Forestry and Food of Slovenia, and Slovenian Research and Innovation Agency (ARIS Programs P4-0431 and P4-0072).

Cold stress induces rapid gene-specific changes in the levels of H3K4me3 and H3K27me3 in Arabidopsis thaliana.

When exposed to low temperatures, plants undergo a drastic reprogramming of their transcriptome in order to adapt to their new environmental conditions, which primes them for potential freezing temperatures. While the involvement of transcription factors in this process, termed cold acclimation, has been deeply investigated, the potential contribution of chromatin regulation remains largely unclear. A large proportion of cold-inducible genes carries the repressive mark histone 3 lysine 27 trimethylation (H3K27me3), which has been hypothesized as maintaining them in a silenced state in the absence of stress, but which would need to be removed or counteracted upon stress perception. However, the fate of H3K27me3 during cold exposure has not been studied genome-wide. In this study, we offer an epigenome profiling of H3K27me3 and its antagonistic active mark H3K4me3 during short-term cold exposure. Both chromatin marks undergo rapid redistribution upon cold exposure, however, the gene sets undergoing H3K4me3 or H3K27me3 differential methylation are distinct, refuting the simplistic idea that gene activation relies on a switch from an H3K27me3 repressed chromatin to an active form enriched in H3K4me3. Coupling the ChIP-seq experiments with transcriptome profiling reveals that differential histone methylation only weakly correlates with changes in expression. Interestingly, only a subset of cold-regulated genes lose H3K27me3 during their induction, indicating that H3K27me3 is not an obstacle to transcriptional activation. In the H3K27me3 methyltransferase curly leaf (clf) mutant, many cold regulated genes display reduced H3K27me3 levels but their transcriptional activity is not altered prior or during a cold exposure, suggesting that H3K27me3 may serve a more intricate role in the cold response than simply repressing the cold-inducible genes in naïve conditions.

The TELOMERE REPEAT BINDING proteins TRB4 and TRB5 function as transcriptional activators of PRC2-controlled genes to regulate plant development

Plant-specific transcriptional regulators called TELOMERE REPEAT BINDING proteins (TRBs) combine two DNA-binding domains, the GH1 domain, which binds to linker DNA and is shared with H1 histones, and the Myb/SANT domain, which specifically recognizes the telobox DNA-binding site motif. TRB1, TRB2, and TRB3 proteins recruit Polycomb group complex 2 (PRC2) to deposit H3K27me3 and JMJ14 to remove H3K4me3 at gene promoters containing telobox motifs to repress transcription. Here, we demonstrate that TRB4 and TRB5, two related paralogs belonging to a separate TRB clade conserved in spermatophytes, regulate the transcription of several hundred genes involved in developmental responses to environmental cues. TRB4 binds to several thousand sites in the genome, mainly at transcription start sites and promoter regions of transcriptionally active and H3K4me3-marked genes, but, unlike TRB1, it is not enriched at H3K27me3-marked gene bodies. However, TRB4 can physically interact with the catalytic components of PRC2, SWINGER, and CURLY LEAF (CLF). Unexpectedly, we show that TRB4 and TRB5 are required for distinctive phenotypic traits observed in clf mutant plants and thus function as transcriptional activators of several hundred CLF-controlled genes, including key flowering genes. We further demonstrate that TRB4 shares multiple target genes with TRB1 and physically and genetically interacts with members of both TRB clades. Collectively, these results reveal that TRB proteins engage in both positive and negative interactions with other members of the family to regulate plant development through both PRC2-dependent and -independent mechanisms.

Telo boxes within the AGAMOUS second intron recruit histone 3 lysine 27 methylation to increase petal number in rose (Rosa chinensis) in response to low temperatures.

The petals of rose (Rosa sp.) flowers determine the ornamental and industrial worth of this species. The number of petals in roses was previously shown to be subject to fluctuations in ambient temperature. However, the mechanisms by which rose detects and responds to temperature changes are not entirely understood. In this study, we identified short interstitial telomere motifs (telo boxes) in the second intron of AGAMOUS (RcAG) from China rose (Rosa chinensis) that play an essential role in precise temperature perception. The second intron of RcAG harbors two telo boxes that recruit telomere repeat binding factors (RcTRBs), which interact with CURLY LEAF (RcCLF) to compose a repressor complex. We show that this complex suppresses RcAG expression when plants are subjected to low temperatures via depositing H3K27me3 marks (trimethylation of lysine 27 on histone H3) over the RcAG gene body. This regulatory mechanism explains the low-temperature-dependent decrease in RcAG transcript levels, leading to the production of more petals under these conditions. Our results underscore an interesting intron-mediated regulatory mechanism governing RcAG expression, enabling rose plants to perceive temperature cues and establish petal numbers.

Identification of epigenetically regulated genes involved in plant-virus interaction and their role in virus-triggered induced resistance

BackgroundPlant responses to a wide range of stresses are known to be regulated by epigenetic mechanisms. Pathogen-related investigations, particularly against RNA viruses, are however scarce. It has been demonstrated that Arabidopsis thaliana plants defective in some members of the RNA-directed DNA methylation (RdDM) or histone modification pathways presented differential susceptibility to the turnip mosaic virus. In order to identify genes directly targeted by the RdDM-related RNA Polymerase V (POLV) complex and the histone demethylase protein JUMONJI14 (JMJ14) during infection, the transcriptomes of infected mutant and control plants were obtained and integrated with available chromatin occupancy data for various epigenetic proteins and marks.ResultsA comprehensive list of virus-responsive gene candidates to be regulated by the two proteins was obtained. Twelve genes were selected for further characterization, confirming their dynamic regulation during the course of infection. Several epigenetic marks on their promoter sequences were found using in silico data, raising confidence that the identified genes are actually regulated by epigenetic mechanisms. The altered expression of six of these genes in mutants of the methyltransferase gene CURLY LEAF and the histone deacetylase gene HISTONE DEACETYLASE 19 suggests that some virus-responsive genes may be regulated by multiple coordinated epigenetic complexes. A temporally separated multiple plant virus infection experiment in which plants were transiently infected with one virus and then infected by a second one was designed to investigate the possible roles of the identified POLV- and JMJ14-regulated genes in wild-type (WT) plants. Plants that had previously been stimulated with viruses were found to be more resistant to subsequent virus challenge than control plants. Several POLV- and JMJ14-regulated genes were found to be regulated in virus induced resistance in WT plants, with some of them poisoned to be expressed in early infection stages.ConclusionsA set of confident candidate genes directly regulated by the POLV and JMJ14 proteins during virus infection was identified, with indications that some of them may be regulated by multiple epigenetic modules. A subset of these genes may also play a role in the tolerance of WT plants to repeated, intermittent virus infections.

Increased long-distance and homo-trans interactions related to H3K27me3 in Arabidopsis hybrids.

In plants, the genome structure of hybrids changes compared with their parents, but the effects of these changes in hybrids remain elusive. Comparing reciprocal crosses between Col × C24 and C24 × Col in Arabidopsis using high-throughput chromosome conformation capture assay (Hi-C) analysis, we found that hybrid three-dimensional (3D) chromatin organization had more long-distance interactions relative to parents, and this was mainly located in promoter regions and enriched in genes with heterosis-related pathways. The interactions between euchromatin and heterochromatin were increased, and the compartment strength decreased in hybrids. In compartment domain (CD) boundaries, the distal interactions were more in hybrids than their parents. In the hybrids of CURLY LEAF (clf) mutants clfCol × clfC24 and clfC24 × clfCol , the heterosis phenotype was damaged, and the long-distance interactions in hybrids were fewer than in their parents with lower H3K27me3. ChIP-seq data revealed higher levels of H3K27me3 in the region adjacent to the CD boundary and the same interactional homo-trans sites in the wild-type (WT) hybrids, which may have led to more long-distance interactions. In addition, the differentially expressed genes (DEGs) located in the boundaries of CDs and loop regions changed obviously in WT, and the functional enrichment for DEGs was different between WT and clf in the long-distance interactions and loop regions. Our findings may therefore propose a new epigenetic explanation of heterosis in the Arabidopsis hybrids and provide new insights into crop breeding and yield increase.

The SlARF4-SlHB8 regulatory module mediates leaf rolling in tomato

Leaf is the main photosynthetic organ in plants and the primary energy source all along the plant life. Given the beneficial role of leaf rolling in improving photosynthetic efficiency and yield in specific environmental conditions, a better understanding of the factors and molecular mechanisms underlying this process is highly suited. Previously, the SlARF4 knocking out mutant exhibited upward curly leaf showed higher resistance to water deficit which driving us to uncover the function of SlARF4 in regulating the curly leaf formation. In this study, we unraveled the unexplored role of the SlARF4-SlHB8 module of transcription factors in the development of leaf rolling. Both SlARF4 loss-of-function and SlHB8 overexpressing tomato plants exhibited upward-rolled leaves, reflecting the active role of the two genes in controlling leaf rolling. Dual-luciferase reporter assays and phenotypic analysis of hybrid progenies suggested that SlHB8 acts downstream of SlARF4 in curly leaf formation. SlARF4 and SlHB8 influence the development of leaf palisade tissues via modulating the expression of genes associated with curly leaf formation. SEM analysis revealed no significant differences in leaf epidermal cells between the two leaf-rolling mutants and the wild type, indicating that curly leaves of arf4 and SlHB8-OE do not result from the asymmetric leaf epidermal cell growth. Our data provide novel insight into the molecular mechanism of abaxial-adaxial determination involving SlARF4 and SlHB8 and reveals that leaf rolling operates via different regulation mechanisms in tomato and Arabidopsis model plant.

CURLY LEAF modulates apoplast liquid water status in Arabidopsis leaves.

The apoplast of plant leaves, the intercellular space between mesophyll cells, is normally largely filled with air with a minimal amount of liquid water in it, which is essential for key physiological processes such as gas exchange to occur. Phytopathogens exploit virulence factors to induce a water-rich environment, or "water-soaked" area, in the apoplast of the infected leaf tissue to promote disease. We propose that plants evolved a "water soaking" pathway, which normally keeps a nonflooded leaf apoplast for plant growth but is disturbed by microbial pathogens to facilitate infection. Investigation of the "water soaking" pathway and leaf water control mechanisms is a fundamental, yet previously overlooked, aspect of plant physiology. To identify key components in the "water soaking" pathway, we performed a genetic screen to isolate Arabidopsis (Arabidopsis thaliana) severe water soaking (sws) mutants that show liquid water overaccumulation in the leaf under high air humidity, a condition required for visible water soaking. Here, we report the sws1 mutant, which displays rapid water soaking upon high humidity treatment due to a loss-of-function mutation in CURLY LEAF (CLF), encoding a histone methyltransferase in the POLYCOMB REPRESSIVE COMPLEX 2 (PRC2). We found that the sws1 (clf) mutant exhibits enhanced abscisic acid (ABA) levels and stomatal closure, which are indispensable for its water soaking phenotype and mediated by CLF's epigenetic regulation of a group of ABA-associated NAM, ATAF, and CUC (NAC) transcription factor genes, NAC019/055/072. The clf mutant showed weakened immunity, which likely also contributes to the water soaking phenotype. In addition, the clf plant supports a substantially higher level of Pseudomonas syringae pathogen-induced water soaking and bacterial multiplication, in an ABA pathway and NAC019/055/072-dependent manner. Collectively, our study sheds light on an important question in plant biology and demonstrates CLF as a key modulator of leaf liquid water status via epigenetic regulation of the ABA pathway and stomatal movement.

Distribution of Yellow Curly Leaf Disease in Chili Plantations in Southeast Sulawesi and Identification of the Causes

Penyakit daun keriting kuning pada pertanaman cabai di Sulawesi Tenggara telah dilaporkan sejak tahun 2018, yaitu di Kabupaten Kolaka Timur dan Kota Kendari. Gejala penyakit di lapangan semakin meluas seiring dengan perluasan penanaman cabai. Penelitian bertujuan menghitung kembali insidensi penyakit daun keriting kuning pada pertanaman cabai di Sulawesi Tenggara, mengidentifikasi serangga yang berasosiasi dengan tanaman cabai, dan mengidentifikasi penyebab penyakitnya. Pengamatan insidensi penyakit dilakukan di pertanaman cabai yang berada di Kabupaten Bombana, Konawe Selatan, Konawe, Kendari, Kolaka Timur, Kolaka, dan Kolaka Utara. Identifikasi kutukebul dilakukan berdasarkan karakter morfologi. Deteksi dan identifikasi begomovirus menggunakan metode polymerase chain reaction, yang dilanjutkan dengan analisis sikuensing. Rata-rata insidensi penyakit daun keriting kuning di tujuh kabupaten ialah 36%–90%. Spesies kutukebul yang ditemukan adalah Aleurotrachelus trachoides. Fragmen DNA spesifik begomovirus berukuran 580 pb berhasil diamplifikasi dari sampel tanaman cabai asal tujuh kabupaten di Sulawesi Tenggara. Analisis sikuen mengonfirmasi infeksi Pepper yellow leaf curl Indonesia virus pada pertanaman cabai di Kabupaten Kolaka, Kolaka Utara, Bombana, Konawe, dan Konawe Selatan.

Transcriptomic and epigenomic analyses revealed that polycomb repressive complex 2 regulates not only developmental but also stress responsive metabolism in Brassica rapa.

Polycomb group proteins (PcG) play a crucial role in developmental programs in eukaryotic organisms, including plants. PcG-mediated gene repression is achieved by epigenetic histone modification on target chromatins. Loss of PcG components leads to severe developmental defects. CURLY LEAF (CLF), a PcG component in Arabidopsis, catalyzes the trimethylation of histone H3 on lysine 27 (H3K27me3), a repressive histone mark in numerous genes in Arabidopsis. In this study, we isolated a single homolog of Arabidopsis CLF, namely, BrCLF, in Brassica rapa ssp. trilocularis. Transcriptomic analysis revealed that BrCLF participated in B. rapa developmental processes, such as seed dormancy, leaf and flower organ development, and floral transition. BrCLF was also involved in stress signaling and stress-responsive metabolism, such as aliphatic and indolic glucosinolate metabolism in B. rapa. Epigenome analysis showed that H3K27me3 was substantially enriched in genes related to these developmental and stress-responsive processes. Thus, this study provided a basis for elucidating the molecular mechanism of the PcG-mediated regulation of development and stress responses in B. rapa.

Induksi dan Multiplikasi Kalus Eucalyptus sp. pada Berbagai Media Callus Induction Medium (CIM) Secara In Vitro

Callus Induction and Multiplication of Eucalyptus sp. on Various Media Callus Induction Medium (CIM) In Vitro. Callus induction in vitro can be done by planting specific part of plant such as leaf, root, stem, etc in aseptic condition by using medium that contains 2,4-D, a plat regulator for promoting callus formation. One of the objectives of callus production in vitro is to form secondary metabolites, because eucalyptus is known as a producer of essential oils. This research is an initial study for in vitro production of secondary metabolites for further research. The aim of this study was to determine the most suitable callus induction medium for callus formation and callus multiplication for Eucalyptus grown in vitro. This research was designed as a Completely Randomized Design (CRD), five treatments, repeated five times. Woody Plant Medium (WPM) was used as the basic media. Several Callus induction Medium (CIM) were used as treatments, i.e. CIM1 = 1 ppm 2,4-D; CIM2 = 1,5 ppm 2,4-D; CIM3 = 2 ppm 2,4-D; CIM4 = 1 ppm NAA+1 ppm BAP; and CIM5 = 2 ppm NAA+ 2 ppm BAP. Variables observed were the time of curly leaf formation, time of the emerging callus, colour and texture of the callus, callus’s weight, callus surface area, the weight of callus after multiplication. The result showed that CIM4 and CIM5 had the fastest emerging callus (7 days after planting), CIM1 and CIM2 formed green and crumb texture of callus, CIM1 produced the highest of callus’s weight, and CIM2 resulted in the highest of callus’s surface area. In conclusion, the most suitable CIM for callus induction was CIM1, while for callus multiplication was CM2.

CURLY LEAF is required for the auxin-dependent regulation of 3-dimensional growth specification in Physcomitrium patens.

The no gametophores 4 ( nog4-R ) mutant cannot make the transition from 2-dimensional (2D) to 3-dimensional (3D) growth in Physcomitrium patens and forms side branch initials that are largely fated to become sporophyte-like structures. We describe the three different developmental trajectories adopted by the nog4-R mutant, all of which result in indeterminate growth and defects in cell division plane orientation. A candidate gene approach confirmed that the causative mutation resided in the CURLY LEAF gene, and we highlight a previously uncharacterized role for CURLY LEAF in maintaining auxin homeostasis in P. patens .

Testing of Superiority and Resistance of Cayenne Pepper of Blaze 12 Variety to Pepper Yellow Leaf Curl Disease

Cayenne pepper is one of the important vegetables and has high economic value. This plant can be cultivated in various types of soil in the lowlands to the highlands. Chilies are very much liked by Indonesian people because of it is spicy and contains various nutrients and vitamins. The purpose of this study was to study and determine the productivity and resistance to yellow curly leaf disease of Blaze 12 varieties compared to comparison varieties. The demand for chili continues to increase from year to year but chili production is not optimal. Efforts to increase crop production have several obstacles related to seed quality, crop cultivation technique, pest and disease attacks. Pepper yellow leaf curl virus (PYLCV) is a disease that can reduce production and cause crop failure. One of the ways to improve seed quality is through a breeding process to support chili productivity and improve other characters such as resistance to disease.Testing of variety is a final stage in plant breeding before varieties are released and commercialized. The purpose of this study was to study and determine the productivity and resistance to yellow curly leaf disease of Blaze 12 varieties compared to comparison varieties. The results of the analysis variance showed a significant effect on the characters: leaf length, leaf width, fruit length, fruit diameter, skin thickness, weight per fruit, number of fruit per plant and yield per hectare. Resistance to yellow curl leaf disease of the Blaze 12 variety was as same as VP-3 and better than VP-1 and VP-2, which showed resistance with mild symptoms. The character of cayenne pepper yellow curl disease is closely related to leaf area.

DETEKSI MOLEKULER DAN ANALISIS GENETIK BEGOMOVIRUS PADA TANAMAN CABAI DI DESA PEMATANG DONOK

The chili cultivation center in Bengkulu Province, which is located in Pematang Donok Village, Kepahiang Regency, has found many symptoms of yellow curly leaf disease, but there have been no reports of viral species that infect the chili plantations. Sampling was done by purposive sampling method based on the symptoms of chili plants infected with Begomovirus. Virus detection using polymerase chain reaction (PCR) method using general primer Begomovirus (SPG 1/2). The PCR product was sent to First Base, Malaysia for DNA sequencing. Sequencing data were analyzed using MEGA 6 (Molecular Evolutionary Genetics Analysis) software for phylogenetic tree construction. Based on field observations, the incidence of disease ranged from 96.4%-100% and the whitefly vector insect (Bemisia tabaci) was found. PCR using general primers Begomovirus obtained DNA bands measuring 912 bp according to the primary target. Based on the Blastn results, the nucleotide sequences of 3 Begomovirus samples from Pematang Donok Village had nucleotide similarities with the Pepper yellow leaf curl Indonesia virus (PYLCIV) isolates from Bali and Java contained in the GenBank database, with a nucleotide similarity value of 91.77%-98.99%.

TOR represses stress responses through global regulation of H3K27 trimethylation in plants.

Target of rapamycin (TOR) functions as a central sensory hub linking a wide range of external stimuli to gene expression. The mechanisms underlying stimulus-specific transcriptional reprogramming by TOR remain elusive. Here, we describe an in silico analysis in Arabidopsis demonstrating that TOR-repressed genes are associated with either bistable or silent chromatin states. Both states regulated by the TOR signaling pathway are associated with a high level of histone H3K27 trimethylation (H3K27me3) deposited by CURLY LEAF in a specific context with LIKE HETEROCHROMATIN PROTEIN1. The combination of the two epigenetic histone modifications H3K4me3 and H3K27me3 implicates a bistable feature that alternates between an 'on' and an 'off' state, allowing rapid transcriptional changes upon external stimuli. The chromatin remodeler SWI2/SNF2 ATPase BRAHMA activates TOR-repressed genes only at bistable chromatin domains to rapidly induce biotic stress responses. Here, we demonstrate both in silico and in vivo that TOR represses transcriptional stress responses through global maintenance of H3K27me3.

Identification and characterization of a curly-leaf locus CL1 encoding an IAA2 protein in Brassica napus

The leaf is the main organ for rapeseed photosynthesis, and its morphology influences photosynthetic efficiency and supports increased planting density and yield. However, the molecular regulatory mechanism of leaf morphology in Brassica napus is poorly understood, restricting progress in breeding for the trait. We describe a novel dominant mutation, curly leaf 1 (cl1), which confers uneven dorsal–ventral axis development, irregular cellular structure and influenced gravitropic response in the seedling stage. The CL1 locus was mapped to a 1.573-Mb interval on chromosome A05 using simple sequence repeat (SSR) markers, and co-segregated with the phenotype of plants in the curly F2 population. A substitution (P62S) was identified in the highly conserved degron motif (GWSPV) of the IAA2 protein in the cl1 mutant, and the P62S substitution impaired the interaction between IAA2 and TIR1 in the presence of auxin, influencing auxin signaling. The P62S substitution-induced curly leaf phenotype was verified by ectopic expression of BnaA05.iaa2 in Arabidopsis and B. napus. Our findings explain the function of IAA2 in rapeseed, providing a foundation for future investigation of auxin signaling and the mechanisms underlying leaf development in B. napus.