Rice LHS1/OsMADS1 Controls Floret Meristem Specification by Coordinated Regulation of Transcription Factors and Hormone Signaling Pathways

SEPALLATA (SEP) MADS box transcription factors mediate floral development in association with other regulators. Mutants in five rice (Oryza sativa) SEP genes suggest both redundant and unique functions in panicle branching and floret development. LEAFY HULL STERILE1/OsMADS1, from a grass-specific subgroup of LOFSEP genes, is required for specifying a single floret on the spikelet meristem and for floret organ development, but its downstream mechanisms are unknown. Here, key pathways and directly modulated targets of OsMADS1 were deduced from expression analysis after its knockdown and induction in developing florets and by studying its chromatin occupancy at downstream genes. The negative regulation of OsMADS34, another LOFSEP gene, and activation of OsMADS55, a SHORT VEGETATIVE PHASE-like floret meristem identity gene, show its role in facilitating the spikelet-to-floret meristem transition. Direct regulation of other transcription factor genes like OsHB4 (a class III homeodomain Leu zipper member), OsBLH1 (a BEL1-like homeodomain member), OsKANADI2, OsKANADI4, and OsETTIN2 show its role in meristem maintenance, determinacy, and lateral organ development. We found that the OsMADS1 targets OsETTIN1 and OsETTIN2 redundantly ensure carpel differentiation. The multiple effects of OsMADS1 in promoting auxin transport, signaling, and auxin-dependent expression and its direct repression of three cytokinin A-type response regulators show its role in balancing meristem growth, lateral organ differentiation, and determinacy. Overall, we show that OsMADS1 integrates transcriptional and signaling pathways to promote rice floret specification and development.

Phenotypic characters of rice landraces reveal independent lineages of short-grain aromatic indica rice

Rice landraces are lineages developed by farmers through artificial selection during the long-term domestication process. Despite huge potential for crop improvement, they are largely understudied in India. Here, we analyse a suite of phenotypic characters from large numbers of Indian landraces comprised of both aromatic and non-aromatic varieties. Our primary aim was to investigate the major determinants of diversity, the strength of segregation among aromatic and non-aromatic landraces as well as that within aromatic landraces. Using principal component analysis, we found that grain length, width and weight, panicle weight and leaf length have the most substantial contribution. Discriminant analysis can effectively distinguish the majority of aromatic from non-aromatic landraces. More interestingly, within aromatic landraces long-grain traditional Basmati and short-grain non-Basmati aromatics remain morphologically well differentiated. The present research emphasizes the general patterns of phenotypic diversity and finds out the most important characters. It also confirms the existence of very unique short-grain aromatic landraces, perhaps carrying signatures of independent origin of an additional aroma quantitative trait locus in the indica group, unlike introgression of specific alleles of the BADH2 gene from the japonica group as in Basmati. We presume that this parallel origin and evolution of aroma in short-grain indica landraces are linked to the long history of rice domestication that involved inheritance of several traits from Oryza nivara, in addition to O. rufipogon. We conclude with a note that the insights from the phenotypic analysis essentially comprise the first part, which will likely be validated with subsequent molecular analysis.

The polycomb group gene EMF2B is essential for maintenance of floral meristem determinacy in rice

Polycomb Repressive Complex 2 (PRC2) represses the transcriptional activity of target genes through trimethylation of lysine 27 of histone H3. The functions of plant PRC2 have been chiefly described in Arabidopsis, but specific functions in other plant species, especially cereals, are still largely unknown. Here we characterize mutants in the rice EMF2B gene, an ortholog of the Arabidopsis EMBRYONIC FLOWER2 (EMF2) gene. Loss of EMF2B in rice results in complete sterility, and mutant flowers have severe floral organ defects and indeterminacy that resemble loss-of-function mutants in E-function floral organ specification genes. Transcriptome analysis identified the E-function genes OsMADS1, OsMADS6 and OsMADS34 as differentially expressed in the emf2b mutant compared with wild type. OsMADS1 and OsMADS6, known to be required for meristem determinacy in rice, have reduced expression in the emf2b mutant, whereas OsMADS34 which interacts genetically with OsMADS1 was ectopically expressed. Chromatin immunoprecipitation for H3K27me3 followed by quantitative (q)RT-PCR showed that all three genes are presumptive targets of PRC2 in the meristem. Therefore, in rice, and possibly other cereals, PRC2 appears to play a major role in floral meristem determinacy through modulation of the expression of E-function genes.

Characterization of rice small heat shock proteins targeted to different cellular organelles

Small heat shock proteins (sHSPs) are a family of ATP-independent molecular chaperones which prevent cellular protein aggregation by binding to misfolded proteins. sHSPs form large oligomers that undergo drastic rearrangement/dissociation in order to execute their chaperone activity in protecting substrates from stress. Substrate-binding sites on sHSPs have been predominantly mapped on their intrinsically disordered N-terminal arms. This region is highly variable in sequence and length across species, and has been implicated in both oligomer formation and in mediating chaperone activity. Here, we present our results on the functional and structural characterization of five sHSPs in rice, each differing in their subcellular localisation, viz., cytoplasm, nucleus, chloroplast, mitochondria and peroxisome. We performed activity assays and dynamic light scattering studies to highlight differences in the chaperone activity and quaternary assembly of sHSPs targeted to various organelles. By cloning constructs that differ in the length and sequence of the tag in the N-terminal region, we have probed the sensitivity of sHSP oligomer assembly and chaperone activity to the length and amino acid composition of the N-terminus. In particular, we have shown that the incorporation of an N-terminal tag has significant consequences on sHSP quaternary structure.

Functions for rice RFL in vegetative axillary meristem specification and outgrowth

Roles for the transcription factor RFL in rice axillary meristem development were studied. Its regulatory effects on LAX1, CUC1, and OsPIN3 reveal its functions in axillary meristem specification and outgrowth.Axillary meristems (AMs) are secondary shoot meristems whose outgrowth determines plant architecture. In rice, AMs form tillers, and tillering mutants reveal an interplay between transcription factors and the phytohormones auxin and strigolactone as some factors that underpin this developmental process. Previous studies showed that knockdown of the transcription factor gene RFL reduced tillering and caused a very large decrease in panicle branching. Here, the relationship between RFL, AM initiation, and outgrowth was examined. We show that RFL promotes AM specification through its effects on LAX1 and CUC genes, as their expression was modulated on RFL knockdown, on induction of RFL:GR fusion protein, and by a repressive RFL-EAR fusion protein. Further, we report reduced expression of auxin transporter genes OsPIN1 and OsPIN3 in the culm of RFL knockdown transgenic plants. Additionally, subtle change in the spatial pattern of IR4 DR5:GFP auxin reporter was observed, which hints at compromised auxin transport on RFL knockdown. The relationship between RFL, strigolactone signalling, and bud outgrowth was studied by transcript analyses and by the tillering phenotype of transgenic plants knocked down for both RFL and D3. These data suggest indirect RFL-strigolactone links that may affect tillering. Further, we show expression modulation of the auxin transporter gene OsPIN3 upon RFL:GR protein induction and by the repressive RFL-EAR protein. These modified forms of RFL had only indirect effects on OsPIN1. Together, we have found that RFL regulates the LAX1 and CUC genes during AM specification, and positively influences the outgrowth of AMs though its effects on auxin transport.

Fingerprinting environmental conditions and related stress using stable isotopic composition of rice (Oryza sativa L.) grain organic matter

Sea level rise (SLR) is a primary factor responsible for inundation of low-lying coastal regions across the world, which in turn governs the agricultural productivity. In this study, rice (Oryza sativa L.) cultivated seasonally in the Kuttanad Wetland, a SLR prone region on the southwest coast of India, were analysed for oxygen, hydrogen and carbon isotopic ratios (delta O-18, delta H-2 and delta C-13) to distinguish the seasonal environmental conditions prevalent during rice cultivation. The region receives high rainfall during the wet season which promotes large supply of fresh water to the local water bodies via the rivers. In contrast, during the dry season reduced river discharge favours sea water incursion which adversely affects the rice cultivation. The water for rice cultivation is derived from regional water bodies that are characterised by seasonal salinity variation which co-varies with the delta O-18 and delta H-2 values. Rice cultivated during the wet and the dry season bears the isotopic imprints of this water. We explored the utility of a mechanistic model to quantify the contribution of two prominent factors, namely relative humidity and source water composition in governing the seasonal variation in oxygen isotopic composition of rice grain OM. delta C-13 values of rice grain OM were used to deduce the stress level by estimating the intrinsic water use efficiency (WUEi) of the crop during the two seasons. 1.3 times higher WUE, was exhibited by the same genotype during the dry season. The approach can be extended to other low lying coastal agro-ecosystems to infer the growth conditions of cultivated crops and can further be utilised for retrieving paleo-environmental information from well preserved archaeological plant remains. (c) 2015 Elsevier Ltd. All rights reserved.

System requirement report for ‘Level 2’: national management institutions, Fisheries Department in Cambodia

(51 p.)

STREAM country strategy paper: Cambodia

Cambodia is one of the poorest countries in the world; much of its population live in rural areas and many live below the local poverty line. The management of common property aquatic resources is of over-riding importance to food security and sustainable rural development in Cambodia. The key groups of poor people who use aquatic resources as part of their diverse livelihoods portfolios are subsistence fishers, small-scale aquaculture practitioners and aquatic resources collectors. Subsistence fishers access mainly the rivers, lakes and inundated forests in Tonle Sap provinces, the lower Mekong and Bassac regions and the upper part of the Mekong. Small-scale aquaculture and/or the collection of aquatic resources are most important in provinces that are not rich in fisheries resources including Kompong Speu, Ratnakiri, Mondulkiri, Preah Vihear and Ortdar Meanchey. Freshwater capture fisheries probably contribute more to national food security and the national economy in Cambodia than in any other country in the world. (19 p.)

Proceedings of the workshop on conclusions and lessons learnt from the livelihood study. Phnom Penh, Cambodia, 26-27 September 2002

This workshop is to follow-up the study on livelihoods of farmers and fishers in Kandal, Kampong Chhnang and Kratie provinces in Cambodia. It was the fourth workshop, part of an ongoing series of activities that will inform the Community Fisheries Development (CFD) Office of the Department of Fisheries (DOF), of the development of and changes in the fisheries law as it affects poor users of aquatic resources.(41 p.)

An examination of the Community Fisheries Sub-Decree in Cambodia: changes and developments during the drafting process

This study examines the development of the Community Fisheries Sub-Decree. From its promulgation in late 2000 through its draft form in June 20021, this sub-decree has gone through over 25 drafts. Initially, the writing process involved the public intimately: community meetings were conducted, NGOs contributed their knowledge and expertise, and then, with these consultations in mind, the newly-formed Community Fisheries Development Office (CFDO) in the Department of Fisheries (DoF) drafted the sub-decree. Over the following year and a half, the sub-decree went through numerous changes, some of which fundamentally alter the tenor of the legislation. This includes the deletion of some of the primary issues of rural communities, such as what fishing gears they can use and whether they can participate in patrolling the fishing area. While the final form is still pending, there are many questions as to the positive effects that the sub-decree will ultimately have on people’s livelihoods when it is finally approved. This research was undertaken to track the development of the sub-decree to gain a better understanding of how the Cambodian policy-making system functions. Consultations were undertaken with the DoF and CFDO before the research began, both of which agreed to the study. It is hoped that this report will help not only those interested in community fisheries issues, but also those who want to advocate on other Cambodian development issues as well. Due to the sensitive nature of the subject material, and the fact that the sub-decree is still pending, the author has conducted interviews with the express purpose of keeping them confidential. Therefore, no individual will be quoted directly in this study. In addition, many of the English versions of the drafts examined were unofficial translations. The author has made no effort to correct the English in the drafts. Moreover, there are also questions regarding the drafts and their sequence, as no government records were kept of the process or the changes that were made along the way. These have been compiled after the fact. (33 p.)

Reducing non-point DOC and nitrogen exports from rice fields: A pilot study and quantitative survey to determine the effects of different hydrologic management practices : Deliverable 15.3 Final Report

Rice cultivation at any level in the Sacramento–San Joaquin Delta (existing or expanded) compels the need to quantify surface and subsurface loads of dissolved organic carbon (DOC), disinfection byproduct precursors (DBPPs) and nitrogen. This information can be used to develop Best Management Practices (BMPs) to reduce export of these constituents in order to improve drinking water quality. Although rice cultivation in the Delta is relatively limited, several factors outside of this research could contribute to increased rice acreage in the Delta: • Recently developed rice varieties seem more suitable for the Delta climate than earlier varieties which required warmer conditions; • Previous economic analyses (Appendix A.10) suggest rice is more profitable than corn, a dominant land use in the Delta; • Recent studies on wetlands at Twitchell Island suggest rice production can help mitigate oxidative subsidence (Miller et al. 2000); • The different oxidative states that result from flooding in rice as compared to those found in crops that require drained soils may help control crop specific weeds and nematodes when rice is incorporated into a crop rotation; and • Providing flooded conditions during a greater part of the year than other crops may benefit water birds. ... (PDF contains 249 pages)

An analysis of poverty and aquatic resources use - focusing especially on the livelihoods of the poor - in Cambodia

The main objectives of this report, which is based on the current literature and key informant interviews, is to assess and analyse the nature and distribution of poverty and aquatic resources use, focusing especially on the livelihoods of the poor. It describes and reports different ways of measuring poverty that are used in Cambodia and quantifies the diverse nature and geographic distribution of aquatic resources use in Cambodia. (PDF contains 55 pages)

Information access survey: Cambodia

As the Cambodian government begins to work more closely with local aquatic resources managers from poor rural communities, increased attention is being paid to the use of communication strategies and tools. In particular, the newly established Community Fisheries Development Office (CFDO) of the Department of Fisheries (DOF) is seeking mechanisms to share information about aquatic resources co-management practices and the livelihoods of people who depend upon the resources. The aim of this report is to identify and recommend methods of communication that are appropriate to aquatic resources management stakeholders, focusing in particular on poor rural communities. (Pdf contains 51 pages).

Addressing technical, social and economic constraints to rice fish culture in Laos, emphasising women’s involvement: DFID Aquaculture Research Programme Project R6380Cb, final technical report

[PDF contains 83 pages]