Clarkepovlsen5901

The transmembrane receptor kinase family is the largest protein kinase family in Arabidopsis. Many members of this family play critical roles in plant signaling pathways. However, many of these kinases have yet uncharacterized functions and very little is known about the direct substrates of these kinases. We have developed the "ShortPhos" method, an efficient and simple mass spectrometry (MS)-based phosphoproteomics protocol to perform comparative phosphopeptide profiling of knockout mutants of receptor-like kinases. Through this method, we are able to better understand the functional roles of plant kinases in the context of their signaling networks.Owing to their sessile nature, plants have evolved sophisticated sensory mechanisms to respond quickly and precisely to the changing environment. The extracellular stimuli are perceived and integrated by diverse receptors, such as receptor-like protein kinases (RLKs) and receptor-like proteins (RLPs), and then transmitted to the nucleus by complex cellular signaling networks, which play vital roles in biological processes including plant growth, development, reproduction, and stress responses. The posttranslational modifications (PTMs) are important regulators for the diversification of protein functions in plant cell signaling. Protein phosphorylation is an important and well-characterized form of the PTMs, which influences the functions of many receptors and key components in cellular signaling. Protein phosphorylation in plants predominantly occurs on serine (Ser) and threonine (Thr) residues, which is dynamically and reversibly catalyzed by protein kinases and protein phosphatases, respectively. selleck compound In this review, we focus on the function of protein phosphorylation in plant cell signaling, especially plant hormone signaling, and highlight the roles of protein phosphorylation in plant abiotic stress responses.Plants are an important part of nature because as photoautotrophs, they provide a nutrient source for many other living organisms. Due to their sessile nature, to overcome both biotic and abiotic stresses, plants have developed intricate mechanisms for perception of and reaction to these stresses, both on an external level (perception) and on an internal level (reaction). Specific proteins found within cells play crucial roles in stress mitigation by enhancing cellular processes that facilitate the plants survival during the unfavorable conditions. Well before plants are able to synthesize nascent proteins in response to stress, proteins which already exist in the cell can be subjected to an array of posttranslation modifications (PTMs) that permit a rapid response. These activated proteins can, in turn, aid in further stress responses. Different PTMs have different functions in growth and development of plants. Protein phosphorylation, a reversible form of modification has been well elucidated, and its role in signaling cascades is well documented. In this mini-review, we discuss the integration of protein phosphorylation with other components of abiotic stress-responsive pathways including phytohormones and ion homeostasis. Overall, this review demonstrates the high interconnectivity of the stress response system in plants and how readily plants are able to toggle between various signaling pathways in order to survive harsh conditions. Most notably, fluctuations of the cytosolic calcium levels seem to be a linking component of the various signaling pathways.Protein phosphorylation is an important cellular regulatory mechanism affecting the activity, localization, conformation, and interaction of proteins. Protein phosphorylation is catalyzed by kinases, and thus kinases are the enzymes regulating cellular signaling cascades. In the model plant Arabidopsis, 940 genes encode for kinases. The substrate proteins of kinases are phosphorylated at defined sites, which consist of common patterns around the phosphorylation site, known as phosphorylation motifs. The discovery of kinase specificity with a preference of phosphorylation of certain motifs and application of such motifs in deducing signaling cascades helped to reveal underlying regulation mechanisms, and facilitated the prediction of kinase-target pairs. In this mini-review, we took advantage of retrieved data as examples to present the functions of kinase families along with their commonly found phosphorylation motifs from their substrates.Anther culture is an important biotechnological tool for quick recovery of fixed breeding lines with unique gene combinations that might otherwise disappear in the course of an extended series of segregating generations in conventional breeding methods in rice. The haploid microspores in culture or the resultant haploid plants are converted to doubled haploids (homozygotes). Variation in doubled haploid lines from F1 hybrids is due to the recovery of rare gene combinations by single round of recombination following meiosis. Androgenesis in rice is largely species- and genotype-specific. O. glaberrima responds better to anther culture than O. sativa; and japonica sub-group is more responsive to microspore embryogenesis than indica types. The author provides a detailed protocol of the anther culture technique for doubled haploid production in indica rice hybrids amenable for genetic improvement.Anther culture is the most used technique to produce doubled haploid lines in rice. This technique is well developed in a wide range of indica rice genotypes. However, in japonica type, and more specifically, the Mediterranean japonica, the protocols are yet to be optimized. Japonica and indica have different androgenic response, as well as different induction and regeneration rates, albinism ratios and chromosome doubling competence. The step-by-step anther culture protocol presented in this chapter allows to regenerate doubled haploid rice plantlets from anther microspores in 8 months. We also include an in vitro chromosome doubling protocol to induce doubled haploids from haploid plantlets by immersion in a colchicine solution. This chromosome doubling protocol complements the anther culture by taking advantage of the regenerated haploid plantlets.Wide hybridization is one of the haploid-inducing techniques that can accelerate the breeding process. Obtaining new cultivars is crucial to solve the problem of the constantly growing world population and global increase in demand for food, feed and renewable energy under changing environmental conditions. Here, we present a detailed protocol for obtaining oat (Avena sativa L.) doubled haploids (DHs) by pollination with maize (Zea mays L.). After fertilization, not only oat homozygotes, but also oat × maize hybrid zygotes can be formed, and during early embryo development, maize chromosomes are preferentially eliminated, which ultimately results in haploid plant formation. This chapter describes a method to produce oat DHs by crossing oat with maize, covering all steps from crossings to haploid plant regeneration and chromosome doubling.Production of doubled haploids (DHs) by androgenesis is a promising and convenient alternative to traditionally used breeding techniques. Low response of anther culture and strong genotype dependency in the development of embryo-like structures (ELS) was reported for oat (Avena sativa L.). Total homozygosity has been reached in one generation. This chapter describes a step-by-step protocol that can be useful for androgenesis studies and oat DH line production through anther culture.Here, we describe a method of triticale isolated microspore culture for production of doubled haploid plants via androgenesis. We use this method routinely because it is highly efficient and works well on different triticale genotypes. To force microspores into becoming embryogenic, we apply a 21-day cold pretreatment. The shock of cold facilitates redirecting microspores from their predestined pollen developmental program into the androgenesis pathway. Ovaries are included in our culture methods to help with embryogenesis, and the histone deacytelase inhibitor Trichostatin A (TSA) is added to further improve androgenesis and increase our ability to recover green doubled haploid plants.Isolated microspore culture systems have been designed in maize by several groups, mainly from the late 1980s to early 2000s. However, even with optimized protocols, microspore embryogenesis induction has remained very dependent on the genotype in maize, with elite germplasm generally displaying no response or very low response. Yet, these last few years, significant progress has been accomplished in understanding and controlling microspore embryogenesis induction in model dicot and monocot species. This knowledge may be transferred to maize, and isolated microspore culture may gain new interest in this crop, at least for embryogenesis research. The methods we hereby present in detail permit the purification of 3-12 × 105 viable microspores per maize tassel, at the favorable stage for microspore embryogenesis. When cultured in appropriate liquid media, microspores from responsive genotypes give rise to androgenic embryos, which can then be regenerated into fertile doubled haploid plants.The intergeneric hybridization of wheat (Triticum aestivum L.) with maize (Zea mays L.) enables the production of doubled haploids (DHs) of wheat from all wheat hybrids with high efficiencies. Wheat and maize donor plants are raised in environmentally controlled greenhouses until crossing. Before anthesis, wheat spikes are emasculated and then pollinated with maize. Auxin is applied to each individual wheat floret 1 day after pollination. About 2 weeks after crossing, in vitro embryo culture is performed, enabling the regeneration of haploid wheat plantlets after maize chromosome elimination. Haploid plantlets are transferred to the greenhouse and after recovery, their genome is doubled with colchicine. Haploid plantlets can be sampled for DNA extractions and molecular analyses to aid the rapid discard of undesirable plantlets. Doubled haploid plants are raised in a greenhouse until maturity. Seeds of each fertile DH are harvested and often sown the same year. Several cycles of multiplication and evaluation in replicated plot trials and different geographical locations are then done to select the best candidate(s) for varietal registration.Doubled haploid (DH) plant production belongs to modern biotechnology methods of plant breeding. The main advantage of DH plant production methods is the development of genetically homozygous lines in one generation, whilst in conventional breeding programmes, the development of homozygous lines requires more generations. The present chapter describes an efficient protocol for DH plant production in spelt wheat genotypes using in vitro anther culture.The use of doubled haploid lines improves the efficiency of cultivar development and homozygous genotypes can be obtained in one generation, as opposed to conventional line production, which requires several cycles of self-pollination. However, in durum wheat (Triticum turgidum subsp. durum Desf.), the low efficiency of green plant regeneration and the very high frequency of albino plants hinder the application of this technique.We observed the success of using gynogenesis for durum wheat and the significant influence of growing conditions on ovary and callus development, and on plant regeneration. Our results suggested that the cold pretreatment for 2 weeks is efficient for durum wheat. Furthermore, the addition of 2,4-D, vitamins and glutamine, and the use of maltose as sugar source in media improved the ovary regeneration. We describe in this work an efficient method to regenerate green plants from in vitro durum wheat gynogenesis .