International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 3(7), 54-60, July (2014) Int. Res. J. Biological Sci. International Science Congress Association 54 Wheat Genotypes (Triticum aestivum L.) vary widely in their responses of Fertility traits to high Temperature at AnthesisChoudhary Ram Chandra1*, Sharma Nand Kishor, Kumar Mithilesh and Kumar RajeevDepartment of Agricultural Biotechnology & Molecular Biology, Rajendra Agricultural University, Pusa, P.O. Box 848125, Bihar, INDIA National Agri-Food Biotechnology Institute, Mohali, P.O Box 160071, Punjab, INDIAAvailable online at: www.isca.in, www.isca.me Received 13th February 2014, revised 25th March 2014, accepted 19th April 2014Abstract High temperature is a major environmental factor which limits the production and productivity of wheat in most cereals growing areas of the world. Eighteen wheat (Triticum aestivum L.) genotypes were screened for high temperature with respect to four traits under field and the polyhouse as normal and stressed conditions during winter session of 2011-12 at Pusa farm, Rajendra Agricultural University, Bihar, India. Spikelet fertility, number of grains per spike, number of effective tillers per plant and pollen sterility were measured and compared to the controls (without heat stress) and Sonalika as a check. The results showed a significant difference among all of the traits in stress and control conditions. On the basis of heat susceptibility index (HSI) the genotypes, Pusa gold, PBW 343, Raj 3765, HD 2888, F5-995 and K0583 were found relatively heat tolerant. Our results suggest that there are significant differences among genotypes that can be used in breeding for heat tolerance at pre anthesis and post anthesis stages and the development of high yielding wheat varieties. Keywords: Wheat, HSI, High temperature, spikelet fertility, polyhouse. IntroductionWheat (Triticum aestivum L.) isa self-pollinated and one of the most important cereal grain cropof the world. It is very sensitive crop with respect to terminal temperature or post anthesis heat stress. It occupies second ranks in India in terms of total production and cultivated area after rice. Currently, the production of wheat in India is 94.90 million tonnes and an area is 29.90 million hectare with a yield of about 3 tonnes per hectare. Heat stress or high temperature is an abiotic stress which major limit the yield of wheat in arid, semiarid, tropical, and subtropical parts of the world. When a plant is subjected to water stress, high salinity6 andheavy metal such as arsenic7 it affected major physiological processes, plant growth and ultimately leads to reduce crop productivity and production. Crop like sorghum is relatively salt tolerant because it possess several anatomical and morphological features that enable it to survive in salt affected and water limited environments. Terminal heat stress usually affect the crop at several stages of wheat development like, booting, heading, anthesis and ultimately reduction in grain yield, grain size and grain filling duration10 in the most growing areas of the world. Optimum temperature during its reproductive stages is 15\rC, and for each 1\rC above optimum, a 3-4% decrease in yield11,12 during grain filling duration13,14 was recorded in the Mediterranean environment15. Heat stress on or before anthesis decreases crop yield and rate of grain filling duration16,17,18 respectively. The effect of heat stress on yield and yield attributing traits usually depends on the growth stage during which the high temperature was subjected19. The effect of high temperature stress on post anthesis induces several physiological changes in wheat eventually result in smaller size of grain and it may be attributable to decrease ofgrain filling period and grain filling rate or the combined effect of both20. The rate of annual crop yield also changed due to increased concentration of CO, higher temperature and decreased solar radiation upto certain extent21. Due to lack of rain, high temperature and dry soil are favourable condition for some disease in crop like sesame22. Theeffect of higher temperature stress on fertility traits in stress condition is more pronounced then the plant under typical normal condition23. Most studies in common or bread wheat has focused on heat stress on or around anthesis stage. Although the effect of high temperature during the microsporogenesis has been investigated using microscopy and this study was used to detect differences among heat tolerant and heat sensitive genotypes in high temperature stress condition. Raising the production of wheat is a challenge for the future, there is further need to develop high yielding and heat tolerance wheat varieties suitable for different environments of the country. In sugarcane soil microorganism also play important roledetermining crop productivity24 and saline soil are adversely affect metabolic activities in rice25. In warmer areas to enhance the production and productivity in wheat cultivation, new set of varieties having heat tolerance is required. The main aim of this study was to understand the effect of high temperature stress on fertility traits and to identify suitable cultivars will then be used in wheat breeding programmes for the development of heat tolerant germplasm suitable for targeted environment. International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(7), 54-60, July (2014) Int. Res. J. Biological Sci. International Science Congress Association 55 Material and Methods Experimental site and Treatments: The experiment was conducted in the research farm at Rajendra Agricultural University, Bihar, in the Pusa (25.590 N, 85.400 E; 52.18 m elevation) in a clay type of soil during the winter season of 2011-12. The experiments for morphological study of wheat genotypes were conducted in heat stressed and normal conditions to understand the basis of genotypic differences in fertility traits. Genotypes were grown in a three rows of 3.0 m length spaced at 22.5 cm between row and 15 cm within row between plants. Genotypes were sown in plastic pots under open condition and allowed for proper development with protective irrigation until booting and heading stage. The pots were transferred inside polyhouse during anthesis stage followed by creating a heat stress environment and recorded the data at different stages i.e. anthesis, grain filling and maturity. Compound microscopy (Olympus) was used to examine pollen viability after heat stress was applied. Weather: Weekly meteorological data from the date of sowing to harvesting of crop in the year 2011-12 for the temperature (max and min), relative humidity (max and min) and rainfall were recorded with a 12 h photoperiod at night (table 1). Observation of fertility traits: Observation on four quantitative characters was recorded separately under both conditions. In each plot/pot, five randomly selected plants excluding border ones, to record observations. By taking the data from each replication, the mean value for the treatment was computed for analysis. Spikelet fertility: Spikelet fertility was counted on the basis of seed setting at the time of harvest and calculated as follows26. Spikelet fertility (%)=Number of fertile spikelets/Total number of spikelets per spike X 100. Pollen sterility: Pollens or microspores were collected from the lower, middle and upper parts of spikes before anthesis. Pollen sterility was assessed by smearing mature anthers in 2 percent acetocarmine stain. The pollen sterility was calculated as27. Pollen sterility (%) =Number of unstained pollen grains/Total number of pollen grains X 100 Heat susceptibility index: Heat susceptibility index (HSI) was calculated over stress and non-stress environment by using the formula28: HSI = [l-YD/YP]/D Where, YD = mean of genotypes in stress environment, YP = mean of genotypes under non-stress environment, D = 1-[mean YD of all genotypes/mean YP of all genotypes]. Table-1 Weekly meteorological data during crop growth period in winter session of 2011-12 under normal and stressed conditions (From 50 to 17 standard meteorological weeks, where: Max-maximum, Min-minimum) SMW Dates Temperature ( 0 C) Relative Humidity (%) Rainfall (mm) (N) Normal Stressed Normal Stressed Max Min Max Min Max Min Max Min 50 10 th Dec-16 th Dec 18.4 12 23.6 13.1 87 76 89 77 Nil 51 17 th Dec-23rd Dec 20 11.3 25.1 12 91.2 74 92.4 75 Nil 52 24 th Dec-31 st Dec 19.7 9.7 25 10.2 90.7 68 90.9 68.5 Nil 1 1 st Jan-7 th Jan 20.1 12 24.3 13 90.4 78 90.7 78.7 2.5 2 8 th Jan-14 th Jan 18.6 10 26.2 11.3 90 63 91.3 64 3 3 15 th Jan-21 st Jan 21 9.4 26.9 10 88 57 87 58 Nil 4 22 nd Jan-28 th Jan 20.7 6.7 28 7 91 46 91.4 47 Nil 5 29 th Jan-4 th Feb 22.2 6.5 29.1 7.4 89 42 90.3 43.1 Nil 6 5 th Feb-11 th Feb 23.7 9.5 30 10.4 87 46 88.2 47.1 Nil 7 12 th Feb-18 th Feb 24.7 10.5 30.4 11 86 46 87.5 47 Nil 8 19 th Feb-25 th Feb 26.7 10.9 31.6 12.6 85 45 86.1 46 Nil 9 26 th Feb-4 th Mar 26.7 9.9 31.7 11.1 80 37 82 38 Nil 10 5 th Mar-11 th Mar 29.4 11.4 34.6 12 77 45 79.2 46 Nil 11 12 th Mar-18 th Mar 27.5 11.1 35.9 12.3 88 43 89 43.5 0.9 12 19 th Mar-25 th Mar 31.9 14.9 39.8 15.6 85 38 86.1 39 Nil 13 26 th Mar-1 st Apr 35.4 16.3 40.7 17 85 32 86.3 33 Nil 14 2 nd Apr-8 th Apr 35.4 20.1 37.5 21 77 47 78 47 0.2 15 9 th Apr-15 th Apr 32.4 20.3 43.4 21.3 84 40 85 41 1.5 16 16 th Apr-22 nd Apr 37.1 20.6 44.1 21.7 78 34 79 35 Nil 17 23 rd Apr-29 th Apr 38.9 22 45.2 23.1 69 34 70 35 Nil International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(7), 54-60, July (2014) Int. Res. J. Biological Sci. International Science Congress Association 56 Statistical analysis: For statistical analysis the experiments were laid out in randomized block design and completely randomized design under field (without heat stress) and polyhouse (heat stress) conditions, respectively with three replications. The data was statistically evaluated for analysis of variance using SPSS ver. 16.0 for mean, standard errors, and differences in environmental treatments. Results and Discussion The results from the analysis of variance indicated that significant differences among all the studied characters in varying environment indicating the influence of temperature on genotypes and fertility traits (table 2). Further, it was observed that the presence of significant genotypes and environment interaction indicated that genotypes were performing differently in different test environments. Similar results were reported by several researchers29-31. The per se performance analysis of eighteen bread wheat genotypes for grain yield and yield contributing characters were significantly varied among the genotypes for various fertility traits (figure 1 and table 4) and compare to the Sonalika as a check. Among the genotypes Mons ald's, HD 2285, Iepaca rabe, PBW 343, Raj 3765, HD 2888, VL 914 and K0583 in normal and HD 2285, PBW 343 in stressed has more spikelet fertility then the check Sonalika. Only one genotype, AKAW 4008 in normal and F5-995 in a stressed condition produced highest number of grains per spike than check Sonalika. Four genotypes namely, HD 2733, C 306, Kauz/AA/Kauz and HD 2888 on normal and ten genotypes viz. HD 2285, Iepaca rabe, Pusa gold, PBW 343, Kauz/AA/Kauz, Raj 3765, HD 2888, K0583, SAWSN 3010 and Cuo/79/Prulla in a stressed condition produced a significantly higher number of effective tillers than the check Sonalika, increasing trend in the number of effective tillers showed in stressed may be due to temperature, relative humidity and related prevailing weather conditions stressed. Only one genotype, F5-995 in normal and four genotypes viz. Iepaca rabe, AKAW 4008, PBW 343 and VL 914 in a stressed condition showed higher pollen sterility then the check Sonalika. It may be due to high temperature and high relative humidity in stressed to affect the tapetal degeneration during meiosis and loss of water from microspore32. Similar observation has been reported by some earlier researchers33,34,35. Considering most of the fertility traits K0583, Cuo/79/Prulla, PBW 343, Iepaca rabe, HD 2285 and Kauz/AA/Kauz were found promising with good yield potentials. Heat susceptibility index (HSI) analysis was conducted for three characters namely spikelet fertility, number of grains per spike and pollen sterility (table 3), it is the measure of yield stability36 based on minimization of yield loss under stress, compared to non-stress condition rather than on yield contributing traits under dry/stress condition per se37. The genotypes were differentiated into relatively heat tolerant (HSI1) and relatively heat susceptible (HSI&#x-3.3;女1) groups for each trait38. Except HD 2733, all genotypes including Sonalika were relatively heat tolerant and AKAW 4008 was most relatively heat tolerant genotype for spikelet fertility. Remaining one genotype HD 2733 was most relatively heat susceptible genotype. For number of grains per spike the genotypes PBW 343, Pusa gold, Raj 3765, K0583 and HD 2888 were relatively heat tolerant and F5-995 was found most relatively heat tolerant. Remaining genotypes i.e. Mons ald’s, HD 2285, Iepaca rabe, HD 2733, Halna, Kauz/AA/Kauz, Sonalika, SWASN 3010 and Cuo/79/Prulla were categorised as relatively heat susceptible and AKAW 4008 and C 306 were found most relatively heat susceptible. For pollen sterility all genotypes were showing their HSI value below unity, so these genotypes were relatively heat tolerant. Genotype VL 914 and Kauz/AA/Kauz were most relatively heat tolerant while, no genotype which was susceptible to heat. Only six out of 18 genotypes namely, Pusa gold, HD 2888, Raj 3765, PBW 343 F5-995 and K0583 were found to be relatively tolerant to heat stress for all these characters namely spikelet fertility, number of grains per spike, pollen sterility and these results are in agreement with some earlier workers39,40. To characterise crop species phylogenetic and biochemical profiling studies have received considerable attention in recent years41. Table-2 Analysis of variance for design of experiment for fertility traits in wheat Source of variation Environments Spikelet fertility No. of grains per spike No. of effective tillers per plant Pollen sterility Mean sum of Squares Replication Normal 250.51 3.88 1.121 87.696 Stressed 274.19 27.41 1.017 77.573 Treatment Normal 569.212** 110.021** 9.025** 173.682** Stressed 498.675** 96.773** 6.867** 187.304** Error Normal 147.491 13.23 1.478 20.694 Stressed 103.042 13.241 0.617 37.535 ** Significant at 1% and 5% probability level International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(7), 54-60, July (2014) Int. Res. J. Biological Sci. International Science Congress Association 57 Table-3 Heat susceptibility index (HSI) of wheat genotypes for spikelet fertility, number of grains per spike and pollen sterility Sl. No. Genotype Heat Susceptibility Index (HSI) Spikelet fertility No. of grains per spike Pollen sterility 1. Mons ald's 0.75 3.00 0.35 2. HD 2285 0.32 5.82 -0.27 3. Sonalika 0.35 5.07 0.40 4. Iepaca rabe 0.65 1.04 0.14 5 AKAW 4008 -0.11 11.65 -0.42 6. Halna 0.89 6.53 -0.15 7. Pusa gold 0.49 -5.33 0.34 8. PBW 343 0.04 -2.71 0.03 9. HD 2733 1.01 7.82 0.40 10. C 306 0.44 10.01 -0.36 11. Kauz/AA/Kauz 0.14 3.16 -0.76 12. Raj 3765 0.30 -3.15 0.26 13. HD 2888 0.83 -10.13 0.47 14. VL 914 0.60 2.40 -1.02 15. F5-995 0.38 -33.51 0.63 16. K0583 0.73 -6.61 0.23 17. SWASN 3010 0.57 2.24 -0.03 18. Cuo/79/Prulla 0.36 2.62 0.29 Table-4 Means, coefficient of variance and standard error of 18 genotypes of wheat, evaluated at RAU, Bihar (India) over two environmental conditions Sl. No. Characters Environment Mean CV % Se(m) 1. Spikelet fertility Normal 70.91 17.12 7.01 Stressed 47.01 21.77 16.88 2. No. of grains per spike Normal 44.52 8.17 2.10 Stressed 43.46 8.37 6.05 3. No. of effective tillers per plant Normal 5.55 21.90 0.70 Stressed 5.45 21.90 1.31 4. Pollen sterility Normal 20.93 21.72 2.62 Stressed 19.34 31.67 10.19 Conclusion The present study emphasizes that the mean square of genotypes was significant for all the characters studied indicating significant differences exist among the genotypes. Most of the wheat genotypes were more affected when exposed to heat stress during at the time of anthesis. Analysis of variance revealed highly significant differences among the genotypes for the studied all parameters. Considering all the parameters it may be concluded that relatively stable genotypes may be evaluated at various agro climatic regions for grain yield, tolerance to heat along with other contributing characters and can be used as selection criteria in breeding programmes. 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