10 de mayo de 2023

Meeting notes: "At the forefront of plant research 2023" (III)

This last session is shared with the next meeting "Genomics-assisted breeding for boosting crop and livestock improvement".

Wednesday 10 may

Imagen
https://twitter.com/CRAGENOMICA/status/1656225214785568768

Cristobal Uauy  ‘Crop breeding as a DNA-assembly problem‘

Pictures breeding as creating mosaics genomes from genotypes that bring in traits of interest. Refers to the work of Brinton et al (https://www.nature.com/articles/s42003-020-01413-2) to build haplotypes. SNP chips fail to tell apart pangenomes entries as they target mostly nearly identical regions; instead haplotypes reconstruct the history of Watkins landraces accurately. They also found that landraces can improve the yield of elite cultivars such as Paragon, although with awful plant architectures.

They are also reconstructing haplotypes from raw reads in a scalable way. The goal is to find regions identical by state (IBS). Their window-based K-mer approach is https://github.com/Uauy-Lab/IBSpy; they use read of the reference genome as negative control. In their WGA benchmarks they find that 1SNP in 5Kbp ~ identical by state. 30% of their A-genome data have a divergence that you would expect for wild wheat. In fact they confirmed that T. monococcum had 1% IBS regions with hexaploidy wheat, which are evidence of introgressions. They have also found introgressions of T. timopheevii. Work at U Nottingham suggests any region of the hexploid genome is susceptible to introgression. The method works with barley and maize inbred lines.

They use affinity propagation clustering to simplify haplotypes. They obtain robust haplotypes by comparing K-mers to all pangenome assemblies. If several genotypes have the same pattern against the same reference, they probably are IBS in that region.

Up to 55% of modern cultivars can be traced back to <10 Watkins landraces. In the UK they realized the need to go back crossing to landraces to improve yield.

 

Nils Stein  ‘The barley pan-genome: large structural genome variation is not rare’

Starts by explaining how genomes of barley have powered applications from SNP calling up to genomic selection.  He then explains how 22K barleys were analyzed by GBS and intersected to core collections to pick n=20 genotypes, including one wild barley. He shows alternative ways to depict and summarize pangenomes, mentioning that graphs seem to work well with human genomes, but current tools don’t work well with barley (ie pggb). They had to mask geneless regions and build single-copy gene clusters.

They are now working towards a second version of the pangenome adding 23 wild barley and reaching a total of 76 genotypes, not yet T2T assemblies (talking with Sanger for that). This set seem to plateau for domesticated barleys, not yet for wild ones.  

 

Bruno Contreras-Moreira ‘Upgrading the gene annotation at the population level reveals the diverse pan-gene set of Asian rice’

My talk, a related preprint can be found at: https://www.biorxiv.org/content/10.1101/2023.01.03.520531v1

Code, documentation and examples at: https://github.com/Ensembl/plant-scripts

I paste here the abstract: Oryza sativa and Arabidopsis thaliana are the best sources of gene function information among plant genomes, probably for their role as model species. In rice, two independent efforts (RAP-DB and MSU - the latter no longer updated) curated the Nipponbare genes that researchers refer to in their papers. However, these two gene sets are different, and more importantly, do not include gene models found in other rice cultivars. To address these limitations, our consortium  produced high quality genome assemblies for 15 cultivars representative of all cultivated Asian rice, plus the reference genome (IRGSP), which were then annotated using the same protocol and gene expression data. The goal was to produce a consistent catalog of gene models that could be used by rice breeders around the world and distributed by key resources such as Ensembl Plants, Gramene and UniProt. A software prototype was developed to find collinear genes annotated in the rice set, producing a total of 84,530 gene clusters. Of these,  26,357 were found in 15+ rices (soft-core). We then confirmed that the soft-core set i) contains 94.6% of all the BUSCO protein domains of order Poales, ii) 89% of genes of agronomic value curated by RAP-DB and iii) has the largest support from protein mass spectrometry experiments. Further inspection of collinear genes revealed a large degree of diversity in terms of gene boundaries and a significant number of missing genes and potential loss of function alleles. This work highlights the challenges of defining a consensus gene annotation for a crop when different cultivars and populations are considered.

Dani Zamir  ‘Epistasis time’

Epistasis is the surprise you get when you combine two genes and get something you did not expect, not additive. Based on his recent tomato work with two interacting QTLs in different tomato  chromosomes, where they found significant heterosis for 10% of normal water input (https://www.pnas.org/doi/abs/10.1073/pnas.2205787119) he pushed us to move to pairs of markers, triplets of markers instead of one at a time. 

 

Danelle Seymour  ‘Integrating scales to traverse the genotype-phenotype divide in citrus’

[Scion = injerto] She does a pangenome survey of NLR gene models across Citrus species in the context of the HLB insect-transmited bacterial disease. Then she moves to explain their field trials in FL, US, along 3 years. They also do automatic phenotyping of several traits by taking 30 images per fruit.  

 

 

9 de mayo de 2023

Meeting notes: "At the forefront of plant research 2023" (II)

 Tuesday 09 may

Xuemei Chen ‘microRNAs as signals in cell-cell communication’

She talks about microRNA moving across cells through plasmodesmata. Shows published work unveiling miRNAs produced in shoots that end up in the root, such as https://pubmed.ncbi.nlm.nih.gov/34707241.  The first part of  the talk is mostly about their work on discovering how a particular miRNA moves across cell layers in the root and how its activity is controlled by a protein (KTN1) that mediates its loading into AGO1. The second part is about the role of miR156 in thermomorphogenesis in Arabidopsis thaliana, a process where PIF4 plays a signaling role. She terminates by saying her lab is recruiting postdocs.

 

Stephen P. Long ‘More productive future-proofed crops via manipulation of photosynthesis to address global food security in 2050’

Light conversion efficiency is the least optimized aspect of plant productivity during the green revolution (https://pubmed.ncbi.nlm.nih.gov/18374559), and likely the only one with room for improvement. For two decades the have modelled the photosynthesis as diff equation digital twin. By numerical integration they are looking for bottlenecks. They are testing future 2060 conditions to test their predictions at https://soyface.illinois.edu, finding that increases in [CO2] increase yield at the cost on spending more water (due to open stomata), but with large variability among cultivars. They use tobacco to optimize photosynthesis focusing on three subprocesses (1yr from transformation to field trial, see https://www.annualreviews.org/doi/abs/10.1146/annurev-arplant-070221-024745). All three interventions improved efficiency. Engineered, transgenic cowpea seeds are already being sawn in Nigeria and Puerto Rico. They are now doing multi-location field trials.

 

Cristina Ferrándiz ‘Don’t stop me now: the end of reproduction in annual plants‘

Fruits feedback to flowering buds to arrest flowering at the right time (arrest ~bud dormancy according to RNAseq). Fruits export auxin, which probably has a role, as well as cytokinins (https://pubmed.ncbi.nlm.nih.gov/34963064) and ABA (https://www.nature.com/articles/s41467-018-03067-5). They show the potential of mutants in gene FUL to boost fruit production in A. thaliana and pea (two seasons, two locations in Canada; in other trials with elite bg it had a + effect one season, - the next one).

 

Daniel Van Damme ‘Mechanistic insight into plant endocytosis ‘

Endocytosis plays a role in shutting down membrane proteins that help communicate with the outside world.  Jotnarlogs are genes/proteins absent in (human/yeast) species models that remain hidden until found in other orgs (https://doi.org/10.1016/j.cub.2020.03.068). They work with a TPLATE mutant which is viable but thermosensible and aggregates at 35C, and also with the nosh mutant (lacks a SH3 domain used to identify the cargo, such as ubiquitenable protein BRI1). Recent work investigate the role of disorder regions in other EH-domain proteins that take part in vesicles and recognize specific lipids and bind to clathrin (https://www.biorxiv.org/content/10.1101/2022.03.17.484738v2).

Rosa Lozano-Durán ‘Chasing geminiviruses to understand plant biology’

The most divergent protein in geminiviruses is C4, due to positive selection and its role in pathogenicity [symptom determinant]. It has a cp transit peptide and is also found in other membranes. By following the routes of C4 she has found a plant defense mechanism against virus infection mediated by BAM1 RLK  and links the membrane and the chloroplast (https://www.pnas.org/doi/full/10.1073/pnas.1715556115).   https://www.biorxiv.org/content/10.1101/2022.12.16.520777v1

 

Cosmocaixa has a great science museum

 

Joy Bergelson ‘Field GWAS and experimental evolution experiments reveal genetic tradeoffs in response to temporally and spatially variable selection’

Describes 2yr of common garden experiments in 4 locations (2N, 2S) in Sweden. In 75% S experiments, N lines did best. In N sites no difference, but S lines had less overwinter survival. They also do “evolution plots” where the find that viability selectio > fecundity selection. After GWAS analysis, they conclude that fitness is complex, due to a large number of minor effect loci, with no major candidate. They did observe a “home effect”, which they measure by computing freq(SNPs) home and away and then calculating the skewness of their effect on fitness. They used this approach to short list SNPs for local adaptation, with modest success apparantely.

 

Olivier Hamant ‘How transcriptional noise and mechanical conflicts contribute to organ shape reproducibility‘

Mechanical fluctuation drives cell/organ development (https://doi.org/10.1073/pnas.2008895117). Cell microtubules are good sensors of stress levels. Somehow mechanical tensions is reflected in gene expression.

The same as fluctuation is the key word of the climate reports of IBCC. For breeding it means we should aim at robustness, not only efficiency. Stephen Long argues they are not incompatible. Some ideas: mixed varieties by default (in France about 40% of wheat, not sure in which areas?, seed companies are already selling in France mixes apparently), understand hardiness, is biofuel really the way to go? Not much sense, precision agriculture? Very fragile, best “imprecision agriculture”, participatory research. He invites papers on these ideas for the journal https://www.cambridge.org/core/journals/quantitative-plant-biology

 

Liam Dolan ‘De novo development of polarity in plant cells’

Polarity is inherited after cell division in Marchantia.  How does polaritity appear in a non-polar cell? Their current model is that initially the nucleous is at the center, a few hours later it moves to a pole pulled by actin  microfilaments. The division plate seems to be oriented by blue light, not gravity. This is impaired by phototropin mutants, which encode a protein that accumulates in the membrane.

Meeting notes: "At the forefront of plant research 2023" (I)

While my colleagues @EEAD-CSIC attended a meeting on cereal breeding at Lleida, today I travelled to Barcelona to learn at the meeting 'At the forefront of plant research 2023'. This is organizaded by CRAG and the chosen venue is the CosmoCaixa up in the hill. I will be sharing my notes here.

Jump to the other days here: II, III

Monday 08 may

Britt Koskella ‘Leveraging the above ground (phyllosphere) microbiome for plant healt’

She talks about her work on phytosphere (aereal part) microbiome on tomato. They have found that tomato seeds contain a relatively conserved microbiome composition. When grown in controlled conditions in the greenhouse, plants seem to have really little bacteria. The seed bacteria can hel control the growth of Pseudomonas syringae.

In field experiments they observe that there is transmission from neighbors (https://pubmed.ncbi.nlm.nih.gov/35022514), in fact it is more important than the actual genotype of the plants.Further experiments show that drought induces a loss of bacterial/mycorr richness, targeting specific taxa. 

 

Teva Vernoux ‘Auxin in development: where to respond, when to respond and how to respond?’

Shows a nice picture of shoot apical meristem to describe the parts where class A ARFs triggered to establish  a spatial pattern. Permissive chromative marks surround the gene bodies; the repressive ones cover mainly the gene body. In protoplasts the observe that there’s a large network of TFs repressing ARFs. Using DAPseq & protoplast data they find that combinations of class A,B,C ARFS bind inverted repeats and direct repeats with spacers 8,13 and 5,15 respectively. Their model is that different tuples ARF,CRE have different roles.

 

Marja Timmermans ‘How to make a flat leaf: Pre-patterns, small RNA morphogens and Turing dynamics’

Describes a n=6 TF/miRNAs network that control how a flat leaf is made, 3 expressed adaxial and 3 abaxial (https://pubmed.ncbi.nlm.nih.gov/35318449). There are also two opposing gradients of miRNAs (https://www.cell.com/developmental-cell/pdfExtended/S1534-5807(17)30816-X) which control the translation of the TFs and control leaf polarity by defining robust developmental boundaries.

She and collabs explored Turing models / cell automata, which combine a short-range + loop and long-range – feedback loop, to create leafs and managed to bring it down to n=3 while maintaining the known n=6 interactions and adding a few new ones. Their best model predicts that AS2 has to be a mobile TF.  She alsi mentions https://www.science.org/doi/10.1126/science.aay5433

 

Keith Slotkin ‘Targeted Transposition in Arabidopsis’

His lab works on target site integration. Context: HR has so low freq is unusable in plants. Homology-Directed Repais has medium precision. Another option is NEEJ, with low precision. For these reasons they are exploring using guided-transposition using the Pong/mPing system, a PIF/Harbinger element from rice, that have 1-order magnitude success rate of targeted integration when shipping with a long linker to full ds-cutting Cas9 genes.

What about precision of insertion? 0-7bp (mostly 1-4) changes at the junctions of insertion sites. Ping transposase is used to excise, Cas9 to cut new destination site. Offsites? Yes, apparently not using Cas9 but the transposase instead, which is still viable. So far mPing can  integrate 430-1563bp cargos. They are currently testing it on soybean. There’s a preprint at https://assets.researchsquare.com/files/rs-2679086/v1_covered.pdf?c=1679325102

 

Angela Hancock ‘Adaptation to novel environments’

Walks us along the use of Arabidopsis thaliana projects on population genomics, first in Europe and more recently in Africa, including Cabo Verde (https://www.arabidopsis.org/servlets/TairObject?type=species_variant&id=98), where the growing season is very short (2-3m). They have studied the island (Santo Antao, Fogo) populations and found out that they represent a single migration from N Africa. CV populations flower rapidly to produce more seed (46% variance from a nonsynon mutation in FRIGIDA, see https://www.nature.com/articles/s41467-022-28800-z).

Plants from volcanic Fogo shows chlorosis in standard pots. Ionome experiments show those line have rebalanced Fe/Mn to adapt to their native soil by harboring two mutated transporters with different affinities, one of them a loss of function (https://www.science.org/doi/10.1126/sciadv.abm9385), the other a tandem duplication. These are nice examples of evolutionary innovations.

 

 Hao Yu ‘RNA modification underlies plant development and stress responses’

There are over 170 chemical modifications to RNA, 3UTR m6A (N-methyladenosin) being the most common in A. thaliana as measured with nanopore direct RNA sequencing. He shows examples of roles of m6A modification during development (seed onset, proliferation of SAM, male meiosis [rice]). In the second part of his talk he focus on the rm1 mutant (disordered protein region) and the role of m6a in drought survival, possibly in the ABA range of action. They find that RM1 recruits m6A mRNA to stress granules.

 

Elena Monte ‘Photosignaling for plant adaptation to environmental challenges’

Two stories: rhythmic responses, cotyledon expansion, two processes regulated by PIF/phytochrome. She talks about the role of PIFs + basal ABA in closing stomate during the night and opening at dawn, a process that depends on blue light intensity. They found KAT1,a s K++ channel in guard cells, responds both to ABA (to close) and PIFs (to open at dawn, ChIP peaks upstream). https://www.biorxiv.org/content/10.1101/2023.01.14.524044v1