A Simple and Rapid System for DNA and RNA Isolation from Diverse Plants Using Handmade Kit

Masoomi‐Aladizgeh, Farhad; Jabbari, Leila; Nekouei, Reza Khayam; Aalami, Ali

doi:10.1038/protex.2016.015

Cited by 13 publications

(10 citation statements)

References 5 publications

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“…Total RNA was isolated from approximately 15-50 mg of tissues, using a CTAB method optimized by Masoomi-Aladizgeh et al (2016), followed by applying a RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Briefly, 500 μl of the extraction buffer in addition to 25 μl of beta-mercaptoethanol were added to the ground pollen, and the samples were incubated at 65°C for 5 min.…”

Section: Rna Isolationmentioning

confidence: 99%

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate downregulation as a feature of thermotolerance

et al. 2021

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Reproductive performance in plants is impaired as maximum temperatures consistently approach 40°C. However, the timing of heatwaves critically affects their impact. We studied the molecular responses during pollen maturation in cotton to investigate the vulnerability to high temperature. Tetrads (TEs), uninucleate and binucleate microspores, and mature pollen were subjected to SWATH-MS and RNA-seq analyses after exposure to 38/28°C (day/night) for 5 days. The results indicated that molecular signatures were downregulated progressively in response to heat during pollen development. This was even more evident in leaves, where three-quarters of differentially changed proteins decreased in abundance during heat. Functional analysis showed that translation of genes increased in TEs after exposure to heat; however, the reverse pattern was observed in mature pollen and leaves. For example, proteins involved in transport were highly abundant in TEs whereas in later stages of pollen formation and leaves, heat suppressed synthesis of proteins involved in cell-to-cell communication. Moreover, a large number of heat shock proteins were identified in heat-affected TEs, but these proteins were less abundant in mature pollen and leaves. We speculate that the sensitivity of TE cells to heat is related to high rates of translation targeted to pathways that might not be essential for thermotolerance. Molecular signatures during stages of pollen development after heatwaves could provide markers for future genetic improvement.

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Section: Rna Isolationmentioning

confidence: 99%

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate downregulation as a feature of thermotolerance

et al. 2021

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show abstract

“…Total RNA was isolated from approximately 15-50 mg of pollen developmental stages, including TE, UN, BN and AN, using a CTAB method optimized by Masoomi-Aladizgeh et al, (2016), followed by applying a RNeasy Plant Mini Kit (Qiagen). Briefly, a lysis buffer was prepared consisting of: CTAB (125 mg), EDTA (250 mg), Tris base (625 mg) and NaCl (1250 mg) in 25 ml nuclease-free water.…”

Section: Rna Isolationmentioning

confidence: 99%

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate down-regulation as a feature of thermotolerance

Masoomi‐Aladizgeh

McKay

Asar

et al. 2021

Preprint

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Reproductive performance in plants is impaired as maximum temperatures consistently approach 40°C. However, the timing of heatwaves critically affects their impact. We studied the molecular responses of cotton male reproductive stages, to investigate the vulnerability of maturing pollen to high temperature. Tetrads, uninucleate and binucleate microspores, and mature pollen were subjected to SWATH-MS and RNA-seq analyses after exposure to 38/28°C (day/night) for 5 days. The results indicated that molecular signatures were down-regulated over developmental stages in response to heat. This was more evident in leaves where three-quarters of differentially changed proteins were decreased in abundance. Functional analysis showed that translation of genes increased in tetrads after exposure to heat; however, the reverse pattern was observed in mature pollen and leaves. Proteins involved in transport were highly abundant in tetrads, whereas in later stages of development and leaves, heat suppressed cell-to-cell communication. Moreover, a large number of heat shock proteins (HSPs) were identified in heat-affected tetrads, but these proteins were less abundant in mature pollen and leaves. We speculate that the sensitivity of tetrad cells to heat is related to increased activity of translation involved in non-essential pathways. Molecular signatures during pollen development after heatwaves provide markers for future genetic improvement.

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“…Total nucleic acid (DNA and RNA) from 3-day-old wheat seedlings was extracted following a slightly modified CTAB method [21].…”

Section: Nucleic Acid Extraction and Amplificationmentioning

confidence: 99%

Origin and Distribution of the VRN-A1 Exon 4 and Exon 7 Haplotypes in Domesticated Wheat Species

Muterko

Салина

2018

Agronomy

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Abstract:The high adaptive potential of modern wheat to a wide range of environmental conditions is determined by genetic changes during domestication. Genetic diversity in VRN1 genes is a key contributor to this adaptability. Previously, the association between the transitions C->T within the fourth and seventh exons of VRN-A1, the distinguishing pair haplotypes Ex4C/Ex4T and Ex7C/Ex7T, and the modulation of such agronomically valuable traits as the vernalization requirement duration, frost tolerance and flowering time of wheat have been shown. However, this polymorphism was analyzed in only a few cultivars of Triticum aestivum L., and not in other wheat species. In the present study, VRN-A1 exon 4 and exon 7 were investigated in six tetraploid and five hexaploid wheat species carrying different VRN-A1 alleles. An allele-specific polymerase chain reaction (PCR) assay was optimized to identify the VRN-A1 exon 7 haplotypes. It was found that polymorphism of the VRN-A1 exon 7 originated in wild tetraploid wheat of Triticum dicoccoides Körn, while the mutant exon 4 of this gene originated later in domesticated hexaploid wheat of T. aestivum. Both these polymorphisms are found in all hexaploid wheat species. Analysis of the VRN-A1 exon 4 and exon 7 haplotype combinations found that intact exon 7 and mutant exon 4 are associated with analogous types of exon 4 and 7, respectively. With the exclusion of the Vrn-A1c (IL369) and Vrn-A1j alleles, identified only in hexaploid wheat, all dominant VRN-A1 alleles carry intact exons 4 and 7 (Ex4C/7C haplotype). The Ex4C/4T/7T haplotype was detected in numerous accessions of hexaploid wheat and is associated with the presence of multiple copies of VRN-A1. Overall, modern domesticated hexaploid wheat T. aestivum includes most possible combinations of the VRN-A1 exon 4 and exon 7 haplotypes among polyploid wheat, which are present in different proportions. This contributes to the high adaptive potential to a broad range of environmental conditions and facilitates the widespread distribution of this species throughout the world.

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A Simple and Rapid System for DNA and RNA Isolation from Diverse Plants Using Handmade Kit

Cited by 13 publications

References 5 publications

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate downregulation as a feature of thermotolerance

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate downregulation as a feature of thermotolerance

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate down-regulation as a feature of thermotolerance

Origin and Distribution of the VRN-A1 Exon 4 and Exon 7 Haplotypes in Domesticated Wheat Species

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