Habibeh Zare¹ and Habib Nazem²

¹Department of Biology Payame Noor University (PNU) Iran.

²Department of Biology Shiraz University

Abstract

Sodium chloride at 100 and 200 mM reduced seedling growth in sensitive cultivar more than tolerance one. Ascorbate peroxidase activity in shoots decreased in tolerance but increased in sensitive cultivar.increasing NaCl concentration , first decreased guaicol peroxidase activity in both cultivars but increased afterward.proline content increased at 200 mM NaCl but the differences between the two cultivars were not significant.soluble sugars decreased in both cultivars. Sodium absorption increasing in both cultivars with increase in NaCl and decrease in KCl, but sensitive cultivar absorbed more sodium than tolerant cultivar. In contrast ,under the same conditions ,tolerant cultivar absorbed more potassium than sensitive cultivar.

Keywords

Rap flower;salt stress;tolerance mechanism (proline;Antioxidant enzyme and soluble sugars)

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Copy the following to cite this article: Zare H, Nazem H. Screening of a Number 8 Rape Flower Cultivars Based on the Accumulat Osmoticum to Salt Tolerance. Biomed Pharmacol J 2011;4(1)

Copy the following to cite this URL: Zare H, Nazem H. Screening of a Number 8 Rape Flower Cultivars Based on the Accumulat Osmoticum to Salt Tolerance. Biomed Pharmacol J 2011;4(1). Available from: http://biomedpharmajournal.org/?p=1703

Introduction

Salinity  stress is the most serious of restricting factors in plant performance crops that are sensitive to salinity among plants are sensitive to salt, resistant to the little amount of salt in soil depends on root ability in preventing of arrival harmful ions to leaves because ,the storage of fresh water is supplied from underground water for a great amount, this water is reduced by frequent consumption for drinking and irrigation so,the need to salt and acrid(bitter) water, such as sea water ,is increased. These explaintation, diagnosis and identification of plant cultivars that are resistant to salinity appears more important .in this survey, salinity effects (Nacl)on growth and development and physiological and biochemical responses to salinity stress in canola has been studied. Canola is a name used for rape oil seed. This plant is belong to Brassicaceae

Family (64) vegetative form generally is non-woody, alternative leaves ,cluster flower and regular flowers, this plant has fruits in the form of silique4.

Rape has third rank after soybean and cottonseed . in comparison to soybean that has 20% oil and 40% protein ,canola has 40% oil and 23% protein(75) chemical compounds in rape in each gram, the leaf has 3.83 gr water ,2.9 gr protein ,1.7 gr lipid,11.2 hydrocarbon ,1.8 fiber ,136 mg calcium, 38 mg phosphor 4.6 mg Fe , 0.08 thiamin.

In about medical characteristics ,has been said that seed powder of this plant is used for cancer and its ointment is used to treat chronic coughing (26). Canola oil with low level of saturated fatty oil acids and the great amount of unsaturated fatty oil acids such as oleic acid, in amount of 61 percent, has demonstrated that this oil is effective in reducing cholesterol and harmful lipid in blood(LDL)5. fall and spring types are available . the spring ones  include hayola(3.8),PF,Hayola(401) cultivars . fall ones include ,Talaie ,kelver+and Ecapi. Hayola(308) is one of hybrids that has very large seed. This hybrid has been originated from Canada .rape  is one of the most resistant plants against salinity .special characteristics and its adaptation to weather condition in almost places in country,result in development of this culture become point hope in supplying raw edible oil2. So that its necessary to identify the varieties of rape that is more resistant to salinity , in order to increase this crop.

Material and Methods

The seeds of four cultivars were supplied from agriculture research center in Borazjan. Before   application , the  seeds were disinfected.

By HCLO for 10 minutes and after several washings, were used in different experiments. cultivars used in these experiments , include:

Hayola 401

Hayola 308

Talaie

Preparation of solutions needed in this experiment, we used Nacl solution in concentrations,0,25,50,75,300,200,100(Milli Mollar)

Treatment method: first the seeds were disinfected then 15 uniforms has been placed on disinfected Petri dish on two layers of filter puper. To each Petri dish ,smm. Solution of prepared solutions was added and then dishes contained seeds were placed in windy incubator in 23+/-1 c in dark for 3 days then the number of seeds that are emerged and root length were accounted each treatment unclude,3 repetition and in control group(without Nacl) , we used distilled water.

The effect of HCl on vegetative  growth in seeds. In these experiments, we used perlite  culture with hogland liuid.

As A Base Nutrition Medium

Application Method

Disinfected seeds were distributed evenly on special backet placed in dishes contained water and then were stored in temperature condition in experiment. After emergence , the dishes were transformed to culture room and exposed on florescent light, with 6000 loux intensity and this light period: 16 h in light and 8h in dark.7-day plantlets were transformed to dishes contained perlite +Hogland

The aime of this test,selection of the most resistant and sensitive caltivar to salinity stress.after this test the following tests were done on these (two cultivars) cultivars.

The effects of Macl treatment on Ascorbat peroxide enzyme activity extracted from leaves and root of hayola and talaie’s plantlet.

Necessary material and solutions

In this test we has used 0.1 molar phosphate solution with PH=1 , 10 mili molar EDTA , 2.2 and 22 mili molar  Ascorbat and 0.03 molar H2O2 solution.

Plant preparation

after seed culture we used 14 day plantlet in order to extract peroxide ascorbat enzyme.

The extraction of peroxide ascorbat enzyme first,plantlets were washed by distilled water and dried by facial tissues. Leaves and roots were separated from each other and 150 miligram of root tissues and 250 miligram of leaf tissues were weighted and separately were homogenized. Homogenized solution include,umili litter phosphate buffer 0.1 molar with plt=7 and 0.5 ascorbat solution 22 mm and 0.5 mililit EDTA 90 mili molar.

The reduction of light absorbtion in 290 nm legth wave in 5 second interval for 1 minute by means of spectrophotometer was measured and changes in light absorbtion relative to time were determined.

The effects of treatment NaCl on peroxide gayakol peroxide enzyme activity entracted from plantlet of leaves and root.

Necessary Materials and Solutions

In this experiment ,we used H2O2  0.03  M, phosphate buffer 0.1 Molar ph=7 , gayacol  solution 0.02 Molar and solid kcl.

The method of treatment : in order to measure peroxide gayacol enzyme, increasing light absorbtion in 437 nm wavelength in 15 seconds interval for 5 minutes was recorded and light absorbtion changes relative to time were determined.

“The effects of Nacl on prolin value in two cultivars”

Necessary Materials and Solutions

Nine hydrin solution ,acid acetic ,phosphoric acid, sulfosalicylic acid 3%, toloune and prolin amino acid.

In order to prepare ninehydrin , 60 mililiter acetic acid was added to 2.5 gram of powder form of this and then 20 mililiter of phosphoric acid 6m was added, resulting solution was stored in refrigerator .

The Method of Treatment

Cleaned plantlets were placed in oven for 21 hour ,then , 30 milligram of dried root was weighed and 10 mililiter of sulfo salicylic acid %3 was added . after 24 hours  , 2 milliliter acetic acid and 2 milliliter nine hydrin was added to resulting solution. Then,tubes are placed into warm bath for 1 hour.after cooling ,ycc toloan was added to each tube and was shaked for 20 seconds by vertex set. We could see that two phases were formed.upper phase absorbtion (toloan contained color complex) in 250nm wavelength was measured.

Results

The effect of NaCl on elementary growth in 8 rape cultivars.

Emergence percent and root length in 8 cultivars ,talaie(sensitive) and hayola 308(resistant) was identified.

The effect of Nacl  on emergence in rape  cultivars.

Big letter in each row and small letter in each  column are different according to duncken test has significant difference a=%50

The effect of Nacl on emergence percent in 8 cultivars. In 25 to 100 millimolar Nacl ,hayola cultivar has more emergence  percent than talaie cultivar in  á=%5 have significant reduction relative to control.

The effect of salinity on root length in rape cultivars: results are illustrated in following table. In 25 millimolar NaCl ,root length moderate in all cultivars showed significant reduction in relative to controls. Except, in hayola cultivar that we saw rootlet increasing.reduction in root growth and rootlet length in talaie cultivar was significant(a=%5). In 200 and 300m molar Nacl ,all cultivars showed significant reduction relative to lower concentrations and control groups.

Figure 1: Click here to View figure

 

Figure 2 : Click here to View figure

 

To Lower Concentrations and Control Groups

The comparison of root dry weight in resistant plantlet to salinity : the moderates of  root dry weight in resistant cultivar(h3o8) in 100 and 200 concentration of Nacl relative to control group was reduced and this reduction was not significant (a=%5). In sensitive cultivar (Ta), moderate of root dry weight in 100 and 200 m mol Nacl showed significant reduction relative to control group.

Figure 3 : Click here to View figure

 

Figure 4 : Click here to View figure

The effect of salinity on enzyme activity (peroxide ascorbat) that is extracted from root plantlet of sensitive and resistant cultivar: by increasing in salt concentration , enzyme activity in both cultivar has been decreased. In 100 mmol enzyme activity in H3O8 has not any difference with control group, however, in 200 mmol ,enzyme activity was decreased about 22.5 percent . in talaie cultivar, enzyme activity in 100 and 200 mmol has been decreased about 41.3% and 37.8% in relative to control group (respectively)

Figure 5 : Click here to View figure

 

The effect of salt on enzyme activity of peroxide gayacol extracted from root and leaf of sensitive resistant cultivar. Enzyme activity in leaf, in 100 mmol concentration in both cultivars,showed reduction relative to control group , however , in 200 treatment in hayola cultivar ,enzyme activity has been increased about 14.4%  relative to control group ,and in talaie cultivar, this has been decreased about 8.7% in relative to control group.in root ,in both cultivar (sensitive and resistant) has seen that enzyme activity was increased by concentration increase.in hayolar cultivar,in 200 and 100 treatment ,enzyme activity has been increased about 68% , 38% in comparison to control group (respectively), however in Ta cultivar , in 100 and 200 treatment , enzyme activity has been increased about 10.3% and 20.77%(respectively).

 

Figure 6 : Click here to View figure

 

Figure 7 : Click here to View figure

 

The effect of salinity on prolin value extracted from root of sensitive and resistant cultivar: the amount of root proline in hayola cultivar control group was about 1.47 mmol proline in each gram of root fresh weight. In 200 mmol treatment, this amount was arrived to 19.35 mmol/gram, this means that the amount was increased about 92.4% relative to control group. In talaie cultivar ,control group has shown is m mol/gr prolin in each gram of fresh weight, in 200 mmol treatment the amount of root prolin has been reached to 25 mmo/gr  , this means that we could see an increase about 94% in comparison to control group .

References

1. Bange G.J., Interaction in the potassium and sodium absorption by intact maize seedlings. Plant and Soil. 11: 17-29 (1959).
2. Bernstein L., Crop with and salinity, Van Schiffgaarde, Drainage for agriculture, Agron, Monogr, ASA, Madison, WI 17: 39-54 (1974).
3. Bernstein L, Hayward H.E., Physiology of salt tolerance. Plant Physiol. 9: 25-46 (1958).
4. Binzel M.L., Hess F.D, Bressan R.A, Hasegawa P.M. Intracellular compartmentation of ions in salt adapted tobacco cells. Plant Physiol, 86: 607-614 (1988).
5. Blits K.C., Gallagher J.L., Salinity tolerance of Kosteletzkya virginica. II Root Growth, Lipid content, ion and water relations. Plant Cell and Environment 13: 419-425 (1990).
6. Blumwald E., Engineering salt tolerance in crop plants. University of california, Department of pomology, ISB News Report. (1990).
7. Brugnoli E., Bjorkman O., Growth of cotton under continuous salinity stress: influence on allocation pattern, stomatal and non-stomal components of photosynthesis and dissipation of excess light energy. Planta. 187: 335-347 (1992).
8. Cayley S., Lewis B.A., Guttman H.J., Record MTJR, Characterization of the cytoplasm of Escherichia coli K-12 as a function of external osmolarity. Implications for protein DNA interactions in vivo. Mol Biol 222: 281-300 (1991).
9. Chang H, Siegel BZ, Siegel SM, Salinity induced changes in isoperoxidase in taro, Colocasia esculenta, Phytochem 23: 233-235 (1984).
10. Chauhan RPS, Chauhan CPS and Kumar D., Free proline accumulation in cereals in relation to salt tolerance plant and soil. 57:
    167-175 (1980).
11. Chu TM, Aspinall D, Pleg LG, Stress metabolism, VII salinity and proline accumalation in barley. Aust J. Plant Physiol. 3: 219-299 (1976).
12. Collander R., Selelctive absorption of cations by higher plants. Plant Physiol, 18: 691-720 (1941).
13. Cram WJ., Negative feed back regulation of transport in cells. The maintenance of turgor, volumne and untrient supply. In transport in plants III part A cells. Eds., V luffge and MG pitman. Encyclopedia of plant physicology New series. Springer Verlag Berlin 2A: 284-316 (1976).
14. Csonka LN., Physiological and genetic responses of bacteria to osmotic stress. Microbill Rev. 53: 212-147 (1989).
15. Csonka LN, Hanson AD, Prokaryotic osmoregulation genetics and physiology. A Annu Rev Microbiol. 45: 569-606 (1991).
16. Delanney AJ, Verma DPS, Proline biosynthesis and osmoregulation in plants. Plant J. 4: 215-223 (1993).
17. Duke JA, Brassica napus L., Handbook of Energy Crops. unpublished (1983).
18. Elstner EF, Metabolism of activated oxygen species. The Biotechnology of Plants. 2: 252-315 (1987).
19. Epstein E., The essential role of calcium in selective cation transport by plant cells. Plant physiol. 36: 437-444 (1961).
20. Epstein E Flower TJ, The effect of sodium chloride on enzyme activities from four halophyte species of chenopodiaceae. Phytochemistry 11: 1881-1886 (1972).
21. Fried, M., Oberlander HE, Noggle JC, Kinetics of rubidium absorption and translocation by barley. Plant Physiol 36: 183-191 (1961).
22. Gauch HG, Eaton FM. Effect of saline substrate on hourly levels of carbohydrates and inorganic constituents of barley plants. Plant Physiol. 17: 347-365 (1942).
23. Girija C, Smith BN, Swamy PM, Interactive effects of sodium chloride and calcium chloride on the accumulation of proline and glycinebetaine in peanut (Arachis hypogaea L.) Environmental and Experimental Botany 47(1): 1-10 (2002).
24. Glenn E, Brown JJ, Blumwald E., Salt-tolerance mechanisms and crop potential of halophytes. Critical Reviews in Plant Sciences 18: 227-255 (1999).
25. Golombek SD, Ludders P, Effects of short-term salinity on leaf gas exchange of the fig. (Ficus carica L.,) Plant and Soil. 148: 21-27 (1993).
26. Gossett DR, Millhollon EP, Lucas MC, Antioxidant response to NaCl stress in salt-tolerant and salt-sensitive cultivars of cotton. Crop Sci. 34: 709-714 (1994).
27. Greenway J, Munns R., Mechanisms of salt tolerance in nonhalophytes. Plant Physiol. 31: 149-190 (1980).
28. Greenway H, Setter TL, Accumulation of praline and sucrose during the first hour after transfer of Chlorella emersonii to hight NaCl. Aust J Plant Physiol. 6: 69-79 (1979).
29. Grieve CM, Shannon MC, Dierig DA, Salinity effects on growth, shoot-ion relations, and seed production of Lesquerella fendleri, J. Janick (ed.) pp 239-243 (1999).
30. Hanson AD, Hitz WD, Metabolic responses of mesophytes to plant water deflicits. Plant Physiol. 33: 169-203 (1982).
31. Hanson JB, The function of calcium in plant nutrition. In Advances in Plant Nutrition 1: 149-208 (1984).
32. Harvey DMR, The effects of salinity on ion concentration within the root cells of Zea mays L. Planta 65: 242-248 (1985).
33. Hare PD, Cress WA (Metabolic implications of stress induced proline accumulation in plants. Plant Growth Regulation. 21: 79-102 (1994).
34. Hare PD, Cress WA, Van Staden J., Dissecting the roles of osmolyte accumulation during stress. Plant Cell Envir 21: 535-553 (1998).
35. Harper DB, Harvey BMR, Mechanism of paraquat tolerance in perenuial ryegrass II. Role of superoxide dismutase, catalase and Peroxidas. Plant Cell Environ. 1: 211-215 (1978).