Dataset Information

Peroxidase B6T173 (ZmPrx35) from insect-resistant maize (Zea mays, p84C3) kernels

ABSTRACT: Peroxidase (POD) activity was found to be positively correlated with insect resistance in maize kernels. Thus, we purified active PODs from (Zea mays, p84C3) kernels and subjected to LC-MS/MS analyses. To verify the identified putative plant PODs, we produced them by recombinant expression in E. coli. Whereas B4FFK9 did not display in vitro activity, B6T173 (ZmPrx35) was active and also exhibits a heme cofactor. B6T173 accounts for about 80% of the POD activity in the kernel, according to densitometric evaluation.

INSTRUMENT(S): LTQ

ORGANISM(S): Zea Mays (maize)

SUBMITTER: Robert Winkler

LAB HEAD: Robert Winkler

PROVIDER: PXD002166 | Pride | 2015-08-21

REPOSITORIES: Pride

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Publications

Identification of B6T173 (ZmPrx35) as the prevailing peroxidase in highly insect-resistant maize (Zea mays, p84C3) kernels by activity-directed purification.

López-Castillo Laura M LM López-Arciniega Janet A I JA Guerrero-Rangel Armando A Valdés-Rodríguez Silvia S Brieba Luis G LG García-Lara Silverio S Winkler Robert R

Frontiers in plant science 20150831

Plant peroxidases (PODs) are involved in diverse physiological processes, including defense against pathogens and insects. Contrary to their biological importance, only very few plant PODs have been proven on protein level, because their low abundance makes them difficult to detect in standard proteomics work-flows. A statistically significant positive correlation between POD activity and post-harvest insect resistance has been found for maize (Zea mays, p84C3) kernels. In combining activity-dir ...[more]

PMID: 26379694

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Project description:Maize kernel is an important source of food, feed and industrial raw materials. The illustration of the molecular mechanisms of maize kernel development will be helpful for the manipulation of maize improvements. Although a great many researches based on molecular biology and gecetics have greatly increased our understanding on the kernel development, many of the mechanisms controlling this important process remain elusive. In current study, a microarray with approximately 58,000 probes was used to study the dynamic gene expression during kernel development from the fertilization to physiological maturity. Samples from two consecutive time-points were paired and labeled using different fluorescent dyes (Cy3 and Cy5) and hybridized in the same array. Hybridization of slides was performed according to the manufacturer’s instructions (http://www.maizearray.org/). The hybridized slides were scanned by a Genepix 4000B (Axon, USA). A loop design was applied for running the microarray. Two replicates of each pair of samples were carried out to test both the reproducibility and quality of the chip hybridizations. By comparing six consecutive time-points, namely 1, 5, 10, 15, 25 and 35 days after pollination (DAP), 3,445 differentially expressed genes were identified. These genes were then grouped into 10 clusters showing specific expression patterns using a K-means clustering algorithm. An investigation of function and expression patterns of genes expanded our understanding of the regulation mechanism underlying the important developmental processes, cell division and kernel filling. The differential expression of genes involved in plant hormone signaling pathways suggested that phytohormone might play a critical role in the kernel developmental process. Moreover, regulation of some transcription factors and protein kinases might be involved in the whole developmental process. To obtain the global gene expression profile during maize kernel development, a microarray with approximately 58,000 probes was used. The maize inbred line X178 was planted on the field. Each plant was self-pollinated by hand. The ears were harvested from healthy plants at 1, 5, 10, 15, 25 and 35 days after pollination (DAP), respectively. In order to increase the consistency & uniformity of the isolated kernels, the upper half and about one sixth of ears from the bottom were cut and discarded, the kernels were isolated from the rest part of the ears. Samples at each time-point were collected from at least thirty ears and pooled to represent the line characteristics of X178. Two sub-samples for replication in the microarray analysis were randomly drawn. Samples from two consecutive time-points were paired and labeled using different fluorescent dyes (Cy3 and Cy5) and hybridized in the same array. A loop design was applied for running the microarray. Two replicates of each pair of samples were carried out to test both the reproducibility and quality of the chip hybridizations.

Dataset Information

Peroxidase B6T173 (ZmPrx35) from insect-resistant maize (Zea mays, p84C3) kernels

Publications

Identification of B6T173 (ZmPrx35) as the prevailing peroxidase in highly insect-resistant maize (Zea mays, p84C3) kernels by activity-directed purification.

Similar Datasets

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