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  • Author or Editor: Jiwan Palta x
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In recent years evidence has been presented that implicates the role of free (cytosolic) Ca2+ as a major metabolic and developmental controller in plants. Calcium concentrations in the cytoplasm are kept very low under normal conditions (10-6 to 10-8 M). Small changes in the absolute amount of calcium can create a 10- to 100-fold change in the Ca2+ concentration without upsetting the ionic balance of the ceil. This feature makes Ca2+ an excellent candidate as a second messenger. Thus, a stress induced change in the cytosolic Ca2+ could bring a cellular/plant response to stress. This response is thought to be mediated through activation of Ca2+ and/or Ca2+-calmodulin-dependent protein kinases which in turn mediate the activity of various enzymes via phosphorylation. Recent evidences from the impact of salinity, low temperature, high temperature, and biotic stresses support such a role of calcium. Data on the association between stress-induced injury and perturbation of membrane/cytosolic calcium are available. In addition, evidences for the role of calcium in acclimation to stress have been reported. These studies suggest that manipulation of cellular Ca2+ may be one of the approaches we have on hand to bridge the gap between science and technology.

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Tuberization in potato is known to be under complex biochemical control involving hormones. A number of studies have provided evidence for a critical role of GA in tuberization. There is also evidence that GA in plants can be modulated by a Ca/calmodulin pathway. The purpose of the present study was to determine the influence of supplemental Ca fertilization on tuber size and tuber number. Plantlets of Solanum tuberosum `Russet Burbank' raised in tissue culture were planted in 20-L pots filled with sandy loam field soil with the pH of 6.9 and exchangeable soil Ca level of 350 ppm. All treatments received the same total amount of N (equivalent to the rate of 280 kg·ha-1). Four treatments were evaluated: nonsplit N (from ammonium nitrate), split N (from ammonium nitrate), split N+Ca (from calcium nitrate), split N+Ca (50% N from urea, 50% N from ammonium nitrate and Ca from calcium chloride). The total Ca was applied at the rate equivalent to 168 kg·ha-1 on a split schedule (equally split at four, six, eight and ten weeks after planting). Four months after planting tubers were harvested and evaluated. As expected tuber tissue Ca was increased by Ca application from 144 to 245 μg·g-1. In general, the two Ca treatments had significantly lower tuber number per plant as compared to the nonsplit and split N treatments. A plot of mean tuber Ca and tuber number for individual plants showed a significant negative relationship. Both Ca treatments produced tubers with higher mean tuber weight compared to nonsplit N. This increase in tuber size with Ca application was not apparent when compared with split N treatment. These results show that Ca application to soil can decrease tuber number suggesting that soil Ca may influence tuberization in potato.

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Supplemental calcium application has been shown in our previous work to improve tuber quality and reduce internal defects. We evaluated the response under field conditions of five commerically significant cultivars to a combination of calcium nitrate, calcium chloride and urea (168 kg·ha-1 per season) over three seasons. We were able to determine that the cultivar with the greatest response to supplemental calcium for reduced bruising, `Atlantic' had the lowest levels of tuber tissue calcium. Conversely, cultivars with least response to supplemental calcium, `Dark Red Norland' and `Superior', had the highest levels of tuber tissue calcium. `Snowden' was both intermediate in response to calcium and tuber tissue concentration. Based on data for 3 years, we determined that across cultivars the calcium concentration at which tubers no longer respond is ≈250 ppm and ranges for individual years from 195 to 242 ppm. These results suggest that seasonal variation for individual cultivars may affect the tuber need for calcium for reduced bruising. Although the exact mechanism is not known, we believe that calcium supplemented to bulking tubers may lead to improved cell membrane stability, increased wall structure or enhanced ability of tubers to repair following injury. The results of our study show that supplemental calcium fertilization has the ability to significantly reduce the incidence of tuber bruising for several cultivars.

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Freezing stress resistance is composed of several components namely tolerance, avoidance and acclimation ability. These three components of freezing stress are heritable traits. We have demonstrated that progress in the improvement of freezing stress resistance can be made by individually selecting for various components of this resistance and then recombining them to get the desired plan. Freeze-thaw injury in carrots is manifested as damage to the foliage, cracks on the roots (especially on the crown), and crown root rot. We found that foliage damage following freeze-thaw stress was related to the tolerance of the foliage to ice formation. The formation of cracks in the crown and root tissue was related to formation of ice itself. The carrot breeding lines we tested varied considerably for the crown position in relation to soil surface. The carrot crowns and roots below the soil surface will be better in avoiding ice in the tissue, thus avoiding cracks. The freeze-thaw injury observed on the foliage in the field was highly correlated to the freeze-thaw tolerance of leaf tissue (measured as ion leakage from the leaf tissue) determined by controlled freeze-thaw test in the laboratory. Based on this work we developed a breeding strategy to improve frost hardiness in carrots by combining the characteristics that avoid ice in the crown and root tissues (e.g., crown position underground) with the characteristics that reduce foliage and root injury by ice (freezing tolerance of foliage). By using this strategy we were able to successfully obtain the desired plant. Two hardy carrot hybrids (Eskimo, Artico) were released by Vilmorin and their hardy characteristics have been confirmed under field conditions.

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High temperature effects potato production by reducing overall growth and partitioning of photosynthate to tubers. Recent studies from our laboratory demonstrated that these effects can be reduced by increasing rhizospheric calcium. This present study was conducted to determine if this mitigation of heat stress effect on potato is due to modulation of heat shock protein by calcium during stress. An inert medium and nutrient delivery system capable of maintaining precise rhizospheric calcium levels were used. Biomass was measured and protein samples were collected from potato leaves. Using electroblotting, heat shock proteins were detected by antibodies to Hsp21 and Hsp70 (obtained from Dr. Elizabeth Vierling). Injury by prolonged heat stress was mitigated at calcium concentration >5 ppm. The calcium concentration of leaf and stem tissues were twice as high in 25 ppm calcium-treated plant compared to 1 ppm calcium-treated plants. Total foliage fresh weight was 33% higher and dry weight 20% higher in plants supplied with 25 ppm of calcium than supplied with 1 ppm of calcium. HSP21 was expressed only at high temperature and at greater concentrations in 25 ppm calcium treatment. HSP70 was expressed in both control, 20 °C/15 °C (day/night) and heat-stressed tissue, 35 °C/25 °C (day/night) under various calcium treatments (1 to 25 ppm). Also, there were some differences in HSPs expression patterns between young and mature leaves. Young tissue responded immediately to the heat stress and started to express HSP21 within 1 day. Mature tissue started to express HSP21 after 2 days. HSP21 of young tissue disappeared sooner than mature tissue when heat stress-treated plants were returned to normal conditions. These results support our earlier studies indicating that an increase in rhizospheric calcium mitigate heat stress effects on the potato plant. Furthermore these results suggest that this mitigation may be due to modulation of HSP21by rhizospheric calcium during heat stress.

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Previous studies in our laboratory both in pine needles and potato leaves have shown evidence of an increase in 18: 2 (linoleate) in the purified plasma membrane fraction during cold acclimation. This increase was reversible on deacclimation, thereby suggesting a link between the accumulation of 18: 2 and acquisition of freezing tolerance. These studies suggest that the activity of specific desaturases may be modulated during cold acclimation. This study was aimed at studying the possible involvement of stearoyl-ACP desaturase (delta9) in potato cold acclimation response. Our approach was to study the induction of delta9 desaturase at the transcript level by using potato delta9 desaturase gene specific primers and reverse transcriptase. For this purpose, mRNA from S. tuberosum (cold sensitive, unable to acclimate) and S. commersonii (cold tolerant, able to cold acclimate) was extracted before and after acclimation. Sequence analysis confirmed that the amplified band was delta9 desaturase. Our results show that there is an increase in delta9 desaturase gene transcripts during cold acclimation and that this increase is associated with the cold acclimation response in potato. These results together with previous reports on the increase in 18: 2 in the plasma membrane during cold acclimation give more evidence toward the involvement of stearoyl-ACP desaturase (delta9) in the potato cold response.

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Lipids have been thought to be important largely in membrane structure and energy reserve. It is now evident that lipids and lipid-derived metabolites play a role in many critical cellular processes. Recent studies have shown that membrane lipid-based signaling mediated by phospholipases such as phospholipase A2 (PLA2), phospholipase C (PLC), and phospholipase D (PLD) constitutes a crucial step in plant responses to abiotic and biotic stresses. Phospholipases and their products also play a role during plant growth and development. For example, PLA2-derived lysophospholipids acted as growth regulators that retard senescence of plant tissues. Interestingly, the PLA2 products inhibited the activity of PLD, which has been suggested to be a key enzyme responsible for membrane lipid breakdown leading to plant senescence. Endogenous levels of lysophospholipids, such as lysophosphatidylethanolamine (LPE), could be increased in castor bean leaf discs by the treatment of auxin (50 μM), which is known to be a activator of PLA2. Pretreatment of leaf discs with a PLA2 inhibitor before auxin treatment nullified the auxin effect and rather resulted in accelerated senescence even compared to the nontreated control. Our recent results suggest a potential role of PLA2 products as biologically active molecules mediating hormonal regulation of growth and senescence. One such product LPE is being commercially exploited for retarding senescence and improving shelf life of fruits, vegetables, and cut flowers.

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Ethephon [2-(chloroethyl) phoshonic acid] is used widely to maximize the yield of ripe tomato fruit. However, ethephon causes rapid and extensive defoliation, overripening, and promotes sunscald damage to the fruit. Recent studies from our laboratory have provided evidence that lysophoshatidylethanolamine (LPE) can reduce leaf senescence. We investigated the potential use of LPE to reduce damaging effect of ethephon on tomato foliage. Three-month-old tomato plants (variety Mountain Spring) grown in greenhouse conditions were sprayed with 200 ppm LPE (with 3% ethanol) at 6 and 24 h before ethephon treatment. After 8 days, plants treated with ethephon alone showed about 80% foliar damage while plant treated with LPE before ethephon treatment showed about 25% foliar damage. In a parallel study, LPE together with ethephon was found to maintain three to four times greater chlorophyll content in the leaves compared to ethephon alone. Treatments of LPE did not reduce the fruit ripening response by ethephon. Both sources of LPE were effective in preventing damaging effects of ethephon on the foliage. These results suggest that LPE treatments 6 and 24 h before ethephon application can prevent damaging effects of ethephon on foliage while allowing the acceleration of fruit ripening.

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