Diurnal variation of transitory starch metabolism is regulated by plastid proteins WXR1/WXR3 in Arabidopsis young seedlings

Transitory starch is the portion of starch that is synthesized during the day in the chloroplast and usually used for plant growth overnight. Here, we report altered metabolism of transitory starch in the wxr1/wxr3 (weak auxin response 1/3) mutants of Arabidopsis. WXR1/WXR3 were previously reported to regulate root growth of young seedlings and affect the auxin response mediated by auxin polar transport in Arabidopsis. In this study the wxr1/wxr3 mutants accumulated transitory starch in cotyledon, young leaf, and hypocotyl at the end of night. WXR1/WXR3 expression showed diurnal variation. Grafting experiments indicated that the WXRs in root were necessary for proper starch metabolism and plant growth. We also found that photosynthesis was inhibited and the transcription level of DIN1/DIN6 (Dark-Inducible 1/6) was reduced in wxr1/wxr3. The mutants also showed a defect in the ionic equilibrium of Na+ and K+, consistent with our bioinformatics data that genes related to ionic equilibrium were misregulated in wxr1. Loss of function of WXR1 also resulted in abnormal trafficking of membrane lipids and proteins. This study reveals that the plastid proteins WXR1/WXR3 play important roles in promoting transitory starch degradation for plant growth over night, possibly through regulating ionic equilibrium in the root.

Genome communication in plants mediated by organelle–n­ucleus-located proteins

An increasing number of eukaryotic proteins have been shown to have a dual localization in the DNA-containing organelles, mitochondria and plastids, and/or the nucleus. Regulation of dual targeting and relocation of proteins from organelles to the nucleus offer the most direct means for communication between organelles as well as organelles and nucleus. Most of the mitochondrial proteins of animals have functions in DNA repair and gene expression by modelling of nucleoid architecture and/or chromatin. In plants, such proteins can affect replication and early development. Most plastid proteins with a confirmed or predicted second location in the nucleus are associated with the prokaryotic core RNA polymerase and are required for chloroplast development and light responses. Few plastid–nucleus-located proteins are involved in pathogen defence and cell cycle control. For three proteins, it has been clearly shown that they are first targeted to the organelle and then relocated to the nucleus, i.e. the nucleoid-associated proteins HEMERA and Whirly1 and the stroma-located defence protein NRIP1. Relocation to the nucleus can be experimentally demonstrated by plastid transformation leading to the synthesis of proteins with a tag that enables their detection in the nucleus or by fusions with fluoroproteins in different experimental set-ups. This article is part of the theme issue ‘Retrograde signalling from endosymbiotic organelles’.

Calmodulin is involved in the dual subcellular location of two chloroplast proteins

Cell compartmentalization is an essential process by which eukaryotic cells separate and control biological processes. Although calmodulins are well-known to regulate catalytic properties of their targets, we show here their involvement in the subcellular location of two plant proteins. Both proteins exhibit a dual location, namely in the cytosol in addition to their association to plastids (where they are known to fulfil their role). One of these proteins, ceQORH, a long-chain fatty acid reductase, was analyzed in more detail, and its calmodulin-binding site was identified by specific mutations. Such a mutated form is predominantly targeted to plastids at the expense of its cytosolic location. The second protein, TIC32, was also shown to be dependent on its calmodulin-binding site for retention in the cytosol. Complementary approaches (bimolecular fluorescence complementation and reverse genetics) demonstrated that the calmodulin isoform CAM5 is specifically involved in the retention of ceQORH in the cytosol. This study identifies a new role for calmodulin and sheds new light on the intriguing CaM-binding properties of hundreds of plastid proteins, despite the fact that no CaM or CaM-like proteins were identified in plastids.

Apicomplexan-like parasites are polyphyletic and widely but selectively dependent on cryptic plastid organelles

The phylum Apicomplexa comprises human pathogens such as Plasmodium but is also an under-explored hotspot of evolutionary diversity central to understanding the origins of parasitism and non-photosynthetic plastids. We generated single-cell transcriptomes for all major apicomplexan groups lacking large-scale sequence data. Phylogenetic analysis reveals that apicomplexan-like parasites are polyphyletic and their similar morphologies emerged convergently at least three times. Gregarines and eugregarines are monophyletic, against most expectations, and rhytidocystids and Eleutheroschizon are sister lineages to medically important taxa. Although previously unrecognized, plastids in deep-branching apicomplexans are common, and they contain some of the most divergent and AT-rich genomes ever found. In eugregarines, however, plastids are either abnormally reduced or absent, thus increasing known plastid losses in eukaryotes from two to four. Environmental sequences of ten novel plastid lineages and structural innovations in plastid proteins confirm that plastids in apicomplexans and their relatives are widespread and share a common, photosynthetic origin.

The Light on the Leaves

As noted earlier in the general description of the plant cell, there is a site at which photosynthesis, the process which allows plants to capture sunlight and convert it into energy, occurs. It is this process which has produced oxygen on the planet, food for herbivores, and the cool green hills of Earth we enjoy today. The capture of sunlight allows the grape vine to grow and produce fruit. Of course, while the discussion of the “light reactions” (capture of sunlight) and the subsequent so-called “dark reactions” (producing carbohydrates) is necessarily brief here, it is, nonetheless, an exciting story. We are only now beginning to understand a little of it. The earlier picture (Figure 7.1) of the plant cell is repeated here (Figure 9.1) so that the position of the chloroplast is seen. Refer to page 24 for a discussion of the numbered items. As the leaves begin to develop alongside the apical meristem, proplastids, which are present in the meristematic regions of the plant, are formed. Proplastids grow into plas¬tids (such as amyloplasts and chloroplasts) as they mature in different ways dictated by the plant’s DNA. Some plastids (e.g., chloroplasts) carry pigments, discussed more fully below, that allow them to carry out photosynthesis. Others are used for storage of fat, starch (amyloplasts) or specialized proteins. Still other plastids are used to synthesize specialized compounds needed to form different tissues or to produce compounds for protection (e.g., tannins). Each plastid builds multiple copies of its DNA as it grows. If it is growing rapidly, it makes more genome copies than if it is growing slowly. The genes, ignoring epigenetic (literally “above the gene”) and postgenetic (literally “after the gene”) modifications, about which we still have much to learn, encode plastid proteins, the regulation of whose expression controls differentiation and thus which plastid is eventually formed. However, despite the differentiation of plastids, it appears that many plastids remain connected to each other by tubes called stromules through which proteins can be exchanged.