Expression of a Self-Incompatibility Glycoprotein (S 2 -Ribonuclease) from Nicotiana alata in Transgenic Nicotiana tabacum

Murfett, Jane; Cornish, Edwina C.; Ebert, Paul R.; Bönig, Ingrid; McClure, Bruce; Clarke, Adrienne E.

doi:10.2307/3869475

Cited by 12 publications

(6 citation statements)

References 32 publications

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“…Lycopersicon peruvianum was chosen for these experiments because it is more amenable to transformation than some other solanaceous species such as N. alata (for example see Murfett et al 1992). We first confirmed that, like S-RNase genes in N. alata and P. hybrida, the S 3 -RNase gene of L. peruvianum is also expressed during pollen development.…”

Section: Introductionmentioning

confidence: 60%

“…Interestingly, the S 3 -RNase from transgenic pollen had a slightly higher M r than the stylar protein. Murfett et al (1992) used the 35S promoter to express the N. alata S 2 -RNase in N. tabacum and found no differences in the size or glycosylation pattern of this protein in mature anthers. Similarly, Kirch et al (1995) detected no size difference when they expressed the Solanum tuberosum S 2 -RNase in tobacco pollen.…”

Section: Figmentioning

confidence: 99%

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Pollen-expressed S-RNases are not involved in self-incompatibility in Lycopersicon peruvianum

Dodds

Ferguson

Clarke

et al. 1999

Sexual Plant Reproduction

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Self-incompatibility in solanaceous plants is gametophytically controlled by a multiallelic S-locus.The only known S-locus product is a series of extracellular ribonucleases (the S-RNases) which are expressed in the mature style and determine its self-incompatibility phenotype. Potentially, S-RNases also determine the self-incompatibility phenotype of pollen as some solanaceous plants express this protein for a brief period during anther development. To test this, we first showed that the S 3 -RNase of Lycopersicon peruvianum is expressed during anther development. We then transformed L. peruvianum plants with sense and antisense versions of the S 3 -RNase coding region under the control of a pollen-specific promoter. Pollen from the transgenic plants accumulated S 3 -RNase transcripts and the S 3 -RNase protein was detected immunologically in the sense transgenic plants. However, neither the sense nor the antisense S 3 -RNase constructs altered the self-incompatibility phenotype of pollen from the transgenic plants, demonstrating the S 3 -RNase is not the pollen product of the S 3 -allele.& b d y :

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Section: Introductionmentioning

confidence: 60%

Section: Figmentioning

confidence: 99%

Pollen-expressed S-RNases are not involved in self-incompatibility in Lycopersicon peruvianum

Dodds

Ferguson

Clarke

et al. 1999

Sexual Plant Reproduction

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“…Further extraction of the residual cell material after TBS elution by grinding in liquid nitrogen followed by extraction in a SDS-containing buffer did not recover detectable amounts of TTS proteins. Furthermore, washing isolated transmitting tissue with an isotonic buffer (Murfett et al, 1992), which presumably does not disrupt cells, released the majority of TTS proteins from this tissue ( Figure 4B). These results indicate that in contrast to many extracellular matrix proline-rich proteins that are extensively cross-linked and insoluble (Fry, 1986), TTS proteins are highly soluble, and the majority of these proteins are not cross-linked to the extracellular matrix of the transmitting tissue.…”

Section: Tts Proteins Are Highly Soluble Extracellular Matrix Proteinmentioning

confidence: 99%

“…Tris-buffered saline (TBS; 20 mM Tris-HCI, pH 7.5, 150 mM NaCI) and an isotonic buffer (300 mM sucrose, 150 mM Tris-HCI, pH 8.3, 5 mM EDTA) (Murfett et al, 1992) were used to isolate proteins. In some of the experiments, proteins were eluted from longitudinally bisected tobacco stigma and style in either TBS or the isotonic buffer by gentle swirling (referred to as TBS or isotonic buffer washes).…”

Section: Protein Lsolationmentioning

confidence: 99%

Development and Pollination Regulated Accumulation and Glycosylation of a Stylar Transmitting Tissue-Specific Proline-Rich Protein.

Wang

Cheung

1993

Plant Cell

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The extracellular matrix of stylar transmitting tissues of many angiosperms is enriched in secretory material$ that are believed to be important for interactions with pollen tubes. We have previously characterized two related cDNAs (TTS-1 and TTS-2) for stylar transmitting tissue-specific proline-rich proteins (TTS proteins) from Nicotiana tabacum. We show here that TTS proteins are highly glycosylated proteins with apparent molecular mames ranging between 50 and 100 kD. Results from chemical and enzymatic deglycosylation suggest that TTS proteins have N-linked glycosyl groups, and the extensive glycosylation most probably has resulted from modifications at the proline residues. TTS proteins are localized to the intercellular regions between neighboring transmitting tissue cells, the space in which pollen tubes elongate as they migrate from the stigma toward the ovary. TTS mRNA and protein levels are regulated during pistil development and by pollination. The levels of TTS mRNAs and proteins increase with flower development and reach the maximal levels as flowers approach anthesis. These maximal levels are maintained in the styles for at least 3 to 4 days after pollination, during which time pollen tubes elongate and reach the ovary. Spatially, TTS mRNAs and proteins accumulate first in the stigmatic end of young styles, and their levels progressively increase toward the basal end as pistils mature. Pollination stimulates the levels of TTS mRNAs and proteins in hand-pollinated young styles, which normally accumulate relatively low levels of these TTS gene products. Pollination also qualitatively affects TTS mRNAs and proteins. In pollinated styles, TTS mRNAs are shorter than those in unpollinated styles and underglycosylated TTS protein species begin to accumulate. The elaborate regulatory mechanisms governing TTS mRNAs and proteins during development and by pollination strongly suggest that these proteins may play a functional role in the process of pollination.

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“…The buffers were selected based on their common use and their utility in isolating proteins and other components from the TT. Acetate buffer has been used in the isolation of S 2 -glycoproteins from N. alata styles (Harris et al 1989;Murfett et al 1992); sodium phosphate buffer was used for the extraction of Transmitting Tract-Specific protein and Pistil Extensin-Like Protein III from N. tabacum styles (Cheung et al 1995;Bosch et al 2001Bosch et al , 2003. Tris-HCL was used to extract stigmatic exudates from Papaver rhoaes, glycoprotein from Prunis avium, and arabinogalactan-protein from Gladious (Williams et al 1982;Franklin-Tong et al 1988).…”

Section: Discussionmentioning

confidence: 99%

A novel pollen tube growth assay utilizing a transmitting tract-ablated Nicotiana tabacum style

Eberle¹,

Clasen²,

Anderson³

et al. 2011

Sex Plant Reprod

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Sexual plant reproduction requires multiple pollen-pistil interactions from the stigma (pollen adhesion, hydration, and germination) to the ovary (fertilization). Understanding the factors that regulate pollen tube growth is critical to understanding the processes essential to sexual reproduction. Many pollen tube growth assays (PTGAs) have shorter and slower pollen tube growth when compared to pollen tube growth through the style. The identification and study of factors that regulate pollen tube growth have been impeded by a lack of an efficient and reproducible PTGA. The objective of this research is to develop a robust assay for Nicotiana tabacum pollen tube growth in an environment that supports sustained and normal growth yet is amenable to testing the effects of specific factors. In this paper, we introduce a novel PTGA, which uses pistils from N. tabacum that lack a mature transmitting tract (TT) due to tissue-specific ablation. The TT-ablated style supports normal pollen tube growth and the hollow structure of the style allows modification of the growth environment by direct injection of test material. This PTGA is robust and allows for rapid and accurate measurement of pollen tube length and pollen tube morphology, supporting pollen tube growth from 20 to 35°C and at pH ranging from 4.8 to 7.6. Use of the ablated style for a PTGA is a novel method for the culture of pollen tubes with sustained growth in vivo while permitting the application of treatments to the growing pollen tubes.

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Expression of a Self-Incompatibility Glycoprotein (S 2 -Ribonuclease) from Nicotiana alata in Transgenic Nicotiana tabacum

Cited by 12 publications

References 32 publications

Pollen-expressed S-RNases are not involved in self-incompatibility in Lycopersicon peruvianum

Pollen-expressed S-RNases are not involved in self-incompatibility in Lycopersicon peruvianum

Development and Pollination Regulated Accumulation and Glycosylation of a Stylar Transmitting Tissue-Specific Proline-Rich Protein.

A novel pollen tube growth assay utilizing a transmitting tract-ablated Nicotiana tabacum style

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