Sodium hydrosulfide priming improves the response of photosynthesis to overnight frost and day high light in avocado (Persea americana Mill, cv. 'Hass').

Radiation frost events, which have become more common in the Mediterranean Basin in recent years, inflict extensive damage to tropical/subtropical fruit crops. During radiation frost, sub-zero temperatures are encountered in the dark, followed by high light during the subsequent clear-sky day. One of the key processes affected by these conditions is photosynthesis, which, when significantly inhibited, leads to the enhanced accumulation of reactive oxygen species (ROS) and damage. The use of 'chemical priming' treatments that induce plants' endogenous stress responses is a possible strategy to improve their coping with stress conditions. Herein, we studied the effects of priming with sodium hydrosulfide (NaHS), a donor of hydrogen sulfide (H2 S), on the response of photosynthesis to overnight frost and day high-light conditions in 'Hass' avocado (Persea americana Mill). We found that priming with a single foliar application of NaHS had positive effects on the response of grafted 'Hass' plants. Primed plants exhibited significantly reduced inhibition of CO2 assimilation, a lower accumulation of hydrogen peroxide as well as lower photoinhibition, as compared to untreated plants. The ability to maintain a high CO2 assimilation capacity after the frost was attained on the background of considerable inhibition in stomatal conductance. Thus, it was likely related to the lower accumulation of ROS and photodamage observed in primed 'Hass' plants. This work contributes toward the understanding of the response of photosynthesis in a subtropical crop species to frost conditions and provides a prospect for chemical priming as a potential practice in orchards during cold winters.

Initial proplastid-to-chloroplast differentiation in the developing vegetative shoot apical meristem of Arabidopsis.

In dicot plants, the process by which undifferentiated plastids, termed proplastids, differentiate into mature functional chloroplasts begins in the shoot apical meristem (SAM) and young leaf primordia, and continues along leaf development. In this work, we followed initial chloroplast biogenesis in cells of the SAM in Arabidopsis, during the early stages of germination, using chlorophyll fluorescence as a marker. We found that cells bound to form the SAM in the mature seed embryo lack chlorophyll, while plastids in the rest of the embryo cells are somewhat developed. The initial appearance of chlorophyll in the SAM occurred two days after the onset of germination, was not expedited by higher light intensities, and required a light period of between five to ten hours within these two days. In addition, we found that biogenesis of chloroplasts occurred only in the upper layer of the SAM, as opposed to the two central subtending cell layers. This pattern was maintained, mirroring the developmental status of plastids in the mature vegetative SAM. The work also presents another model for studying proplastid-to-chloroplast development, in which differentiation can be followed in SAM cells in a defined time-wise fashion.

Heterologous expression of Brassica juncea annexin, AnnBj2 confers salt tolerance and ABA insensitivity in transgenic tobacco seedlings.

Annexins are multifunctional proteins with roles in plant development and alleviation of stress tolerance. In the present communication, we report on the effect of heterologous expression of Brassica juncea annexin, AnnBj2 in tobacco. Transgenic tobacco plants expressing AnnBj2 exhibited salt-tolerant and abscisic acid (ABA)-insensitive phenotype at the seedling stage. Biochemical analysis showed that AnnBj2 transgenic plants retained higher chlorophyll and proline content, and lower malondialdehyde (MDA) levels compared to the null line under salt stress. They exhibited better water retention capacity compared to the null segregant (NS) line. AnnBj2 overexpression altered the transcript levels of several stress-related marker genes involved in reactive oxygen species (ROS) scavenging and abiotic stress signaling. Taken together, these results suggest a positive role for AnnBj2 in salt stress response upon heterologous expression in tobacco.

Molecular Approaches for Manipulating Male Sterility and Strategies for Fertility Restoration in Plants.

Usable pollination control systems have proven to be effective system for the development of hybrid crop varieties, which are important for optimal performance over varied environments and years. They also act as a biocontainment to check horizontal transgene flow. In the last two decades, many genetic manipulations involving genes controlling the production of cytotoxic products, conditional male sterility, altering metabolic processes, post-transcriptional gene silencing, RNA editing and chloroplast engineering methods have been used to develop a proper pollination control system. In this review article, we outline the approaches used for generating male sterile plants using an effective pollination control system to highlight the recent progress that occurred in this area. Furthermore, we propose possible future directions for biotechnological improvements that will allow the farmers to buy hybrid seed once for many generations in a cost-effective manner.

Constitutive expression of Brassica juncea annexin, AnnBj2 confers salt tolerance and glucose and ABA insensitivity in mustard transgenic plants.

Annexins belong to a plasma membrane binding (in a calcium dependent manner), multi-gene family of proteins, which play ameliorating roles in biotic and abiotic stresses. The expression of annexin AnnBj2 of Indian mustard is tissue specific with higher expression in roots and under treatments with sodium chloride and abscisic acid (ABA) at seedling stage. The effect of constitutive expression of AnnBj2 in mustard was analyzed in detail. AnnBj2 OE (over expression) plants exhibited insensitivity to ABA, glucose and sodium chloride. The insensitivity/tolerance of the transgenic plants was associated with enhanced total chlorophylls, relative water content, proline, calcium and potassium with reduced thiobarbituric acid reactive substances and sodium ion accumulation. The altered ABA insensitivity of AnnBj2 OE lines is linked to downregulation of ABI4 and ABI5 transcription factors and upregulation of ABA catabolic gene CYP707A2. Furthermore, we found that overexpression of AnnBj2 upregulated the expression of ABA-dependent RAB18 and ABA-independent DREB2B stress marker genes suggesting that the tolerance phenotype exhibited by AnnBj2 OE lines is probably controlled by both ABA-dependent and -independent mechanisms.

Overexpression of Arabidopsis AnnAt8 Alleviates Abiotic Stress in Transgenic Arabidopsis and Tobacco.

Abiotic stress results in massive loss of crop productivity throughout the world. Because of our limited knowledge of the plant defense mechanisms, it is very difficult to exploit the plant genetic resources for manipulation of traits that could benefit multiple stress tolerance in plants. To achieve this, we need a deeper understanding of the plant gene regulatory mechanisms involved in stress responses. Understanding the roles of different members of plant gene families involved in different stress responses, would be a step in this direction. Arabidopsis, which served as a model system for the plant research, is also the most suitable system for the functional characterization of plant gene families. Annexin family in Arabidopsis also is one gene family which has not been fully explored. Eight annexin genes have been reported in the genome of Arabidopsis thaliana. Expression studies of different Arabidopsis annexins revealed their differential regulation under various abiotic stress conditions. AnnAt8 (At5g12380), a member of this family has been shown to exhibit ~433 and ~175 fold increase in transcript levels under NaCl and dehydration stress respectively. To characterize Annexin8 (AnnAt8) further, we have generated transgenic Arabidopsis and tobacco plants constitutively expressing AnnAt8, which were evaluated under different abiotic stress conditions. AnnAt8 overexpressing transgenic plants exhibited higher seed germination rates, better plant growth, and higher chlorophyll retention when compared to wild type plants under abiotic stress treatments. Under stress conditions transgenic plants showed comparatively higher levels of proline and lower levels of malondialdehyde compared to the wild-type plants. Real-Time PCR analyses revealed that the expression of several stress-regulated genes was altered in AnnAt8 over-expressing transgenic tobacco plants, and the enhanced tolerance exhibited by the transgenic plants can be correlated with altered expressions of these stress-regulated genes. Our findings suggest a role for AnnAt8 in enhancing abiotic stress tolerance at different stages of plant growth and development.

Alleviation of methyl viologen-mediated oxidative stress by Brassica juncea annexin-3 in transgenic Arabidopsis.

Plant annexins function as calcium-dependent or -independent phospholipid binding proteins and constitute about 0.1% of total cellular proteins. Some of them were reported to antagonize oxidative stress and protect plant cells. Brassica juncea annexin-3 (AnnBj3) was recently discovered. To gain insight into a possible function of AnnBj3 in oxidative stress response, we investigated the resistance of Arabidopsis thaliana plants expressing AnnBj3 constitutively. Here we report that, AnnBj3 attenuates methyl viologen-mediated oxidative stress in plants. It protected photosynthesis and plasma membrane from methyl viologen-mediated oxidative damage. AnnBj3 detoxifies hydrogen peroxide and showed antioxidative property in vitro. The protein increased total peroxidase activity in transgenics and interfered with other cellular antioxidants, thereby giving an overall cellular protection against methyl viologen-induced cytotoxicity.