Regulation of seed germination in the close Arabidopsis relative Lepidium sativum : a global tissue-specific transcript analysis

The completion of germination in Lepidium sativum and other endospermic seeds (e.g. Arabidopsis [Arabidopsis thaliana]) is regulated by two opposing forces, the growth potential of the radicle (RAD) and the resistance to this growth from the micropylar endosperm cap (CAP) surrounding it. We show by puncture force measurement that the CAP progressively weakens during germination, and we have conducted a time-course transcript analysis of RAD and CAP tissues throughout this process. We have also used specific inhibitors to investigate the importance of transcription, translation, and posttranslation levels of regulation of endosperm weakening in isolated CAPs. Although the impact of inhibiting translation is greater, both transcription and translation are required for the completion of endosperm weakening in the whole seed population. The majority of genes expressed during this process occur in both tissues, but where they are uniquely expressed, or significantly differentially expressed between tissues, this relates to the functions of the RAD as growing tissue and the CAP as a regulator of germination through weakening. More detailed analysis showed that putative orthologs of cell wall-remodeling genes are expressed in a complex manner during CAP weakening, suggesting distinct roles in the RAD and CAP. Expression patterns are also consistent with the CAP being a receptor for environmental signals influencing germination. Inhibitors of the aspartic, serine, and cysteine proteases reduced the number of isolated CAPs in which weakening developed, and inhibition of the 26S proteasome resulted in its complete cessation. This indicates that targeted protein degradation is a major control point for endosperm weakening

Isolation and Characterization of fungal causal agents of black bark disease of apple using the “apple test”

In den letzten, häufig von Trockenheit geprägten Jahren hat sich der Schwarze Rindenbrand an Apfel- und Birnenbäumen in verschiedenen Teilen Deutschlands zunehmend zu einem Problem entwickelt. Ziel der vorliegenden Arbeit war es daher, methodische Grundlagen zur Isolierung der Erreger aus Apfelbäumen und ihrer Charakterisierung im Labor zu erarbeiten. Die Experimente beinhalteten die klassische Isolierung durch Auslegen von erkranktem Gewebe auf Agarmedien sowie ein in der Literatur als „Apfeltest“ beschriebenes Verfahren, bei dem befallene Rinde in Apfelfrüchte gesteckt wird. In der Folge reichern sich Pathogene im Gewebe an und können aus den entstehenden Faulstellen isoliert werden.Aus dem Stammholz eines im Wipfelbereich welkenden Baumes wurden nach Auslegen auf Agarmedium Trametes versicolor, Diaporthe eres sowie Diplodia seriata (bekannt als einer von mehreren Erregern des Schwarzen Rindenbrandes) isoliert. Diese Art wurde auch nach Auslegen befallener Rinde auf Kartoffel-Dextrose-Agar erhalten, ebenso wie Diplodia malorum. Durch Einstecken befallener Rindenstücke in Apfelfrüchte wurden weitere Isolate von D. malorum, drei Isolate von Diplodia bulgarica sowie Lambertella corni-maris, Penicillium sp. und Sclerotinia sclerotiorum gewonnen. Im Biotest mit künstlicher Inokulation mit Reinkulturen riefen die drei letztgenannten Isolate, alle Diplodia-Isolate sowie Diaporthe eres Fruchtfäulen hervor. Die Diplodia-Arten wurden darüber hinaus hinsichtlich der Geschwindigkeit des Hyphenwachstums und der Sporenmorphologie charakterisiert.During the previous years characterized by summer-drought, black canker disease of apple and pear has become a problem in different parts of Germany. The aim of the current work was therefore to develop basic techniques for isolation of the inciting pathogens from apple trees and their characterization in the laboratory. The experiments included the standard isolation of putative pathogens by placing symptomatic tissue on agar media as well as a method described in the literature as „apple test”. For the latter, symptomatic bark pieces are inserted into apple fruits and colonization of the apple tissue is visible as rotting spots. Candidate pathogens can be easily isolated from the colonized apple tissue.After placement of tissue from the trunk of the top of an apple tree expressing wilt symptoms, Trametes versicolor, Diaporthe eres and Diplodia seriata (one of several agents known to cause black canker disease) were isolated. Together with Diplodia malorum, the latter species was also obtained after placement of symptomatic bark pieces on potato dextrose agar. Insertion of diseased bark into apple fruits led to the isolation of further isolates of D. malorum, three isolates of Diplodia bulgarica, Lambertella corni-maris, Penicillium sp. and Sclerotinia sclerotiorum. After inoculation with pure cultures into apple fruits, the latter three, all Diplodia-isolates and Diaporthe eres were pathogenic and caused fruit rots. The Diplodia isolates were further characterised regarding speed of hyphal growth and spore morphology

Jasmonic acid-dependent regulation of seed dormancy following maternal herbivory in Arabidopsis

Maternal experience of abiotic environmental factors such as temperature and light are well known to control seed dormancy in many plant species. Maternal biotic stress alters offspring defence phenotypes, but whether it also affects seed dormancy remains unexplored. We exposed Arabidopsis thaliana plants to herbivory and investigated plasticity in germination and defence phenotypes in their offspring, along with the roles of phytohormone signalling in regulating maternal effects. Maternal herbivory resulted in the accumulation of jasmonic acid-isoleucine and loss of dormancy in seeds of stressed plants. Dormancy was also reduced by engineering seed-specific accumulation of jasmonic acid in transgenic plants. Loss of dormancy was dependent on an intact jasmonate signalling pathway and was associated with increased gibberellin content and reduced abscisic acid sensitivity during germination. Altered dormancy was only observed in the first generation following herbivory, whereas defence priming was maintained for at least two generations. Herbivory generates a jasmonic acid-dependent reduction in seed dormancy, mediated by alteration of gibberellin and abscisic acid signalling. This is a direct maternal effect, operating independently from transgenerational herbivore resistance priming

HRS1 Acts as a Negative Regulator of Abscisic Acid Signaling to Promote Timely Germination of Arabidopsis Seeds

In this work, we conducted functional analysis of Arabidopsis HRS1 gene in order to provide new insights into the mechanisms governing seed germination. Compared with wild type (WT) control, HRS1 knockout mutant (hrs1-1) exhibited significant germination delays on either normal medium or those supplemented with abscisic acid (ABA) or sodium chloride (NaCl), with the magnitude of the delay being substantially larger on the latter media. The hypersensitivity of hrs1-1 germination to ABA and NaCl required ABI3, ABI4 and ABI5, and was aggravated in the double mutant hrs1-1abi1-2 and triple mutant hrs1-1hab1-1abi1-2, indicating that HRS1 acts as a negative regulator of ABA signaling during seed germination. Consistent with this notion, HRS1 expression was found in the embryo axis, and was regulated both temporally and spatially, during seed germination. Further analysis showed that the delay of hrs1-1 germination under normal conditions was associated with reduction in the elongation of the cells located in the lower hypocotyl (LH) and transition zone (TZ) of embryo axis. Interestingly, the germination rate of hrs1-1 was more severely reduced by the inhibitor of cell elongation, and more significantly decreased by the suppressors of plasmalemma H+-ATPase activity, than that of WT control. The plasmalemma H+-ATPase activity in the germinating seeds of hrs1-1 was substantially lower than that exhibited by WT control, and fusicoccin, an activator of this pump, corrected the transient germination delay of hrs1-1. Together, our data suggest that HRS1 may be needed for suppressing ABA signaling in germinating embryo axis, which promotes the timely germination of Arabidopsis seeds probably by facilitating the proper function of plasmalemma H+-ATPase and the efficient elongation of LH and TZ cells

Three endo-β-mannanase genes expressed in the micropylar endosperm and in the radicle influence germination of Arabidopsis thaliana seeds

Mannans are hemicellulosic polysaccharides in the plant primary cell wall (CW). Mature seeds, specially their endosperm cells, have CWs rich in mannan-based polymers that confer a strong mechanical resistance for the radicle protrusion upon germination. The rupture of the seed coat and endosperm are two sequential events during the germination of Arabidopsis thaliana. Endo-β-mannanases (MAN; EC. 3.2.1.78) are hydrolytic enzymes that catalyze cleavage of β1 → 4 bonds in the mannan-polymer. In the genome of Arabidopsis, the endo-β-mannanase (MAN) family is represented by eight members. The expression of these eight MAN genes has been systematically explored in different organs of this plant and only four of them (AtMAN7, AtMAN6, AtMAN2 and AtMAN5) are expressed in the germinating seeds. Moreover, in situ hybridization analysis shows that their transcript accumulation is restricted to the micropylar endosperm and to the radicle and this expression disappears soon after radicle emergence. T-DNA insertion mutants in these genes (K.O. MAN7, K.O. MAN6, K.O. MAN5), except that corresponding to AtMAN2 (K.O. MAN2), germinate later than the wild type (Wt). K.O. MAN6 is the most affected in the germination time course with a t 50 almost double than that of the Wt. These data suggest that AtMAN7, AtMAN5 and specially AtMAN6 are important for the germination of A. thaliana seeds by facilitating the hydrolysis of the mannan-rich endosperm cell walls

The evolution of seeds

The evolution of the seed represents a remarkable life-history transition for photosynthetic organisms. Here, we review the recent literature and historical understanding of how and why seeds evolved. Answering the \u27how\u27 question involves a detailed understanding of the developmental morphology and anatomy of seeds, as well as the genetic programs that determine seed size. We complement this with a special emphasis on the evolution of dormancy, the characteristic of seeds that allows for long \u27distance\u27 time travel. Answering the \u27why\u27 question involves proposed hypotheses of how natural selection has operated to favor the seed life-history phenomenon. The recent flurry of research describing the comparative biology of seeds is discussed. The review will be divided into sections dealing with: (1) the development and anatomy of seeds; (2) the endosperm; (3) dormancy; (4) early seed-like structures and the transition to seeds; and (5) the evolution of seed size (mass). In many cases, a special distinction is made between angiosperm and gymnosperm seeds. Finally, we make some recommendations for future research in seed biology

<i>DELAY OF GERMINATION 1</i> mediates a conserved coat-dormancy mechanism for the temperature- and gibberellin-dependent control of seed germination

Seed germination is an important life-cycle transition because it determines subsequent plant survival and reproductive success. To detect optimal spatiotemporal conditions for germination, seeds act as sophisticated environmental sensors integrating information such as ambient temperature. Here we show that the DELAY OF GERMINATION 1 (DOG1) gene, known for providing dormancy adaptation to distinct environments, determines the optimal temperature for seed germination. By reciprocal gene-swapping experiments between Brassicaceae species we show that the DOG1-mediated dormancy mechanism is conserved. Biomechanical analyses showthat thismechanism regulates the material properties of the endosperm, a seed tissue layer acting as germination barrier to control coat dormancy. We found that DOG1 inhibits the expression of gibberellin (GA)-regulated genes encoding cell-wall remodeling proteins in a temperature-dependent manner. Furthermore we demonstrate that DOG1 causes temperature-dependent alterations in the seed GA metabolism. These alterations in hormone metabolism are brought about by the temperature-dependent differential expression of genes encoding key enzymes of the GA biosynthetic pathway. These effects of DOG1 lead to a temperature-dependent control of endosperm weakening and determine the optimal temperature for germination. The conserved DOG1-mediated coat-dormancymechanismprovides a highly adaptable temperature-sensing mechanism to control the timing of germination.</p

Conserved Roles of CrRLK1L Receptor-Like Kinases in Cell Expansion and Reproduction from Algae to Angiosperms

Receptor-like kinases (RLKs) are regulators of plant development through allowing cells to sense their extracellular environment. They facilitate detection of local endogenous signals, in addition to external biotic and abiotic stimuli. The Catharanthus roseus RLK1-like (CrRLK1L) protein kinase subfamily, which contains FERONIA, plays a central role in regulating fertilization and in cell expansion mechanisms such as cell elongation and tip growth, as well as having indirect links to plant–pathogen interactions. Several components of CrRLK1L signaling pathways have been identified, including an extracellular ligand, coreceptors, and downstream signaling elements. The presence and abundance of the CrRLK1L proteins in the plant kingdom suggest an origin within the Streptophyta lineage, with a notable increase in prevalence in the seeded land plants. Given the function of the sole CrRLK1L protein in a charophycean alga, the possibility of a conserved role in detection and/or regulation of cell wall integrity throughout the Strephtophytes is discussed. Orthologs of signaling pathway components are also present in extant representatives of non-vascular land plants and early vascular land plants including the liverwort Marchantia polymorpha, the moss Physcomitrella patens and the lycophyte Selaginella moellendorffii. Deciphering the roles in development of the CrRLK1L protein kinases in early diverging land plants will provide insights into their ancestral function, furthering our understanding of this diversified subfamily of receptors in higher plants

Peroxidases identified in a subtractive cDNA library approach show tissue-specific transcript abundance and enzyme activity during seed germination of Lepidium sativum

The micropylar endosperm is a major regulator of seed germination in endospermic species, to which the close Brassicaceae relatives Arabidopsis thaliana and Lepidium sativum (cress) belong. Cress seeds are about 20 times larger than the seeds of Arabidopsis. This advantage was used to construct a tissue-specific subtractive cDNA library of transcripts that are up-regulated late in the germination process specifically in the micropylar endosperm of cress seeds. The library showed that a number of transcripts known to be up-regulated late during germination are up-regulated in the micropylar endosperm cap. Detailed germination kinetics of SALK lines carrying insertions in genes present in our library showed that the identified transcripts do indeed play roles during germination. Three peroxidases were present in the library. These peroxidases were identified as orthologues of Arabidopsis AtAPX01, AtPrx16, and AtPrxIIE. The corresponding SALK lines displayed significant germination phenotypes. Their transcripts were quantified in specific cress seed tissues during germination in the presence and absence of ABA and they were found to be regulated in a tissue-specific manner. Peroxidase activity, and particularly its regulation by ABA, also differed between radicles and micropylar endosperm caps. Possible implications of this tissue-specificity are discussed