A novel motif in the NaTrxh N-terminus promotes its secretion, whereas the C-terminus participates in its interaction with S-RNase in vitro.

BACKGROUND: NaTrxh, a thioredoxin type h, shows differential expression between self-incompatible and self-compatible Nicotiana species. NaTrxh interacts in vitro with S-RNase and co-localizes with it in the extracellular matrix of the stylar transmitting tissue. NaTrxh contains N- and C-terminal extensions, a feature shared by thioredoxin h proteins of subgroup 2. To ascertain the function of these extensions in NaTrxh secretion and protein-protein interaction, we performed a deletion analysis on NaTrxh and fused the resulting variants to GFP. RESULTS: We found an internal domain in the N-terminal extension, called Nß, that is essential for NaTrxh secretion but is not hydrophobic, a canonical feature of a signal peptide. The lack of hydrophobicity as well as the location of the secretion signal within the NaTrxh primary structure, suggest an unorthodox secretion route for NaTrxh. Notably, we found that the fusion protein NaTrxh-GFP(KDEL) is retained in the endoplasmic reticulum and that treatment of NaTrxh-GFP-expressing cells with Brefeldin A leads to its retention in the Golgi, which indicates that NaTrxh uses, to some extent, the endoplasmic reticulum and Golgi apparatus for secretion. Furthermore, we found that Nß contributes to NaTrxh tertiary structure stabilization and that the C-terminus functions in the protein-protein interaction with S-RNase. CONCLUSIONS: The extensions contained in NaTrxh sequence have specific functions on the protein. While the C-terminus directly participates in protein-protein interaction, particularly on its interaction with S-RNase in vitro; the N-terminal extension contains two structurally different motifs: Nα and Nß. Nß, the inner domain (Ala-17 to Pro-27), is essential and enough to target NaTrxh towards the apoplast. Interestingly, when it was fused to GFP, this protein was also found in the cell wall of the onion cells. Although the biochemical features of the N-terminus suggested a non-classical secretion pathway, our results provided evidence that NaTrxh at least uses the endoplasmic reticulum, Golgi apparatus and also vesicles for secretion. Therefore, the Nß domain sequence is suggested to be a novel signal peptide.

Conostegia xalapensis (Melastomataceae): an aluminum accumulator plant.

In acidic soils, an excess of Al³âº is toxic to most plants. The Melastomataceae family includes Al-accumulator genera that tolerate high Al³âº by accumulating it in their tissues. Conostegia xalapensis is a common shrub in Mexico and Central America colonizing mainly disturbed areas. Here, we determined whether C. xalapensis is an Al accumulator, and whether it has internal tolerance mechanisms to Al. Soil samples collected from two pastures in the state of Veracruz, Mexico, had low pH and high Al³âº concentrations along with low Ca²âº levels. Leaves of C. xalapensis from pastures showed up to 19,000 mg Al kg⁻¹ DW (dry weight). In laboratory experiments, 8-month-old seedlings treated with 0.5 and 1.0 mM AlCl3 for 24 days showed higher number of lateral roots and biomass. Pyrocatechol violet and hematoxylin staining evidenced that Al localized in epidermis and mesophyll cells in leaves and in epidermis and vascular pith in roots. Scanning electron microscope-energy dispersive X-ray microanalysis of Al-treated leaves corroborated that Al is in abaxial and adaxial epidermis and in mesophyll cells (31.2%) in 1.0 mM Al-treatment. Roots of Al-treated plants had glutathione reductase (EC 1.6.4.2) and superoxide dismutase (EC 1.15.1.1) activity higher, and low levels of O2*⁻ and H2O2. C. xalapensis is an Al-accumulator plant that can grow in acidic soils with higher Al³âº concentrations, and can be considered as an indicator species for soils with potential Al toxicity.

A novel thioredoxin h is secreted in Nicotiana alata and reduces S-RNase in vitro.

Thioredoxins type h are classified into three subgroups. The subgroup II includes thioredoxins containing an N-terminal extension, the role of which is still unclear. Although thioredoxin secretion has been observed in animal cells, there is no evidence suggesting that any thioredoxin h is secreted in plants. In this study, we report that a thioredoxin h, subgroup II, from Nicotiana alata (NaTrxh) is secreted into the extracellular matrix of the stylar transmitting tract tissue. Fractionation studies showed that NaTrxh is extracted along with well characterized secretion proteins such as S-RNases and NaTTS (N. alata transmitting tissue-specific protein). Moreover, an NaTrxh-green fluorescent fusion protein transiently expressed in Nicotiana benthamiana and Arabidopsis thaliana leaves was also secreted, showing that NaTrxh has the required information for its secretion. We performed reduction assays in vitro to identify potential extracellular targets of NaTrxh. We found that S-RNase is one of the several potential substrates of the NaTrxh in the extracellular matrix. In addition, we proved by affinity chromatography that NaTrxh specifically interacts with S-RNase. Our findings showed that NaTrxh is a new thioredoxin h in Nicotiana that is secreted as well as in animal systems. Because NaTrxh is localized in the extracellular matrix of the stylar transmitting tract and its specific interaction with S-RNase to reduce it in vitro, we suggest that this thioredoxin h may be involved either in general pollen-pistil interaction processes or particularly in S-RNase-based self-incompatibility.

Compatibilidad sexual entre dos tipos de Hylocerus (Cactaceae) / Sexual compatibility between two types of Hylocerus (Cactaceae)

There are two types of pitahaya that are cultivated in Yucatan Peninsula of Mexico. They differ mainly in the skin color of the fruit, one of them has a red skin (Uqroo1), while the other has a light yellow skin (Uqroo2) both belong to Hylocereus undatus (Haworth) Britt. & Rose. The yellow skin pitahaya is the sweetest. The studies were conducted using the methods of self pollination and cross pollination, with direct crosses and reciprocal pollination in order to evaluate the effect related to the quantity of fruits that accomplish their growing process. Some characteristic parameters of fruits, thus obtained, were used to detect the possible xenic effect. The control samples were collected from naturally pollinated flowers. The experimental design used was completely random and the experiment was repeated fifteen times. The results revealed that the Uqroo1 turned out to be self-compatible, and their fruits reached one hundred percent of development. On the other hand, Uqroo2 was self-incompatible, that is, it did not accept self pollen. The control samples of both types reached the same weight, and the fruits of the Uqroo2 produced by cross pollination had the same or more weight than the control samples. Several parameters were positively correlated. The coefficient between the number of seeds and the fruit weight was r = 0.84. The highest correlation that was found (r = 0.97) in the fruit weight and the pulp weight within both types. The fruits obtained by cross-pollination maintained the characteristic of the female progenitor. It is suggest intercalate the yellow pitahaya plants with another types or species of pitahaya, other than the Uqroo1.