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EVALUATION OF MORPHO-PHYSIOLOGICAL TRAITS OF TURKISH RICE GENOTYPES IN RESPONSE TO SALT STRESS UNDER IN VITRO CONDITIONS.

Byline: H. Gungor, Y. Cikili and Z. Dumlupinar

Keywords: GGE biplot, ion contents, photosynthetic pigments, rice, salinity stress.

INTRODUCTION

A major portion of global population consumes rice (Oryza sativa L.) at least once a day (Chauhan et al., 2017). Rice was grown on an area of 116.056 ha in Turkey and yielded 920.000 tons of paddy during the year 2016 (TUIK, 2017). Weeds, rice blast disease (Pyricularia oryzae) and salinity are the major factors that limit the rice growth and yield. It is predicted that salinity will cause far more losses to rice yield in the future more than the other factors. Salinity, which restricts crop production, can occur as a result of natural factors as well as improper agricultural practices. Furthermore, most of the soils in rice areas are naturally saline (Hussain et al., 2018). The rice plant is described as moderately salt tolerant (Xiong et al., 2014; Rahman et al., 2016). Therefore, it is an appropriate crop for saline soils (Hussain et al., 2018), but the level of salinity tolerance varies in rice genotypes (Khan et al., 1997).

The salinity affects seed germination by creating osmotic stress due to reduced water uptake or through ionic imbalance due to toxic effects of sodium (Na +) and chloride (Cl-) ions (Hosseini et al., 2003; Hussain et al., 2018). The increased salt level of the conditions lowered the water uptake of the seeds and germination rate (Akbarimoghaddam et al., 2011; Shereen et al., 2011; Balkan et al., 2015; Dogan and Carpici, 2016). Many studies have reported that high level of salinity in rice reduces shoot and root length and shoot and root fresh-dry weight (Haq et al., 2009; Kazemi and Eskandari, 2011; Hussain et al., 2013; Balkan et al., 2015; Solangi et al., 2015; Zafar et al., 2015; Dolo et al., 2016). It is also reported that lower Na, higher K and K/Na ratio provides salt tolerance in rice (Tatar and Gevrek, 2007; Tatar et al., 2010; Hussain et al., 2013; Zafar et al., 2015). The salt stress limits the plant growth besides other abiotic stresses.

The closed stomata, limited CO2 uptake and insufficient photosynthesis are the reasons for the reduction in plant growth as a result of salinity. Singh et al. (2009) reported that salt stress shortened the plant height of rice, reduced the leaf area, lowered the dry weight and limited the photosynthesis and chlorophyll content. In recent years, GGE (genotype, genotype x environmet) biplot analyses were used to evaluate many crop plants in different experiments. One of the main reasons for researchers to use this analysis that it enables to build graphics indicating more than one traits of the genotypes and provides a comparasion between genotypes and traits, since the salt tolerance is highly related with germination and seedling stage traits (Yan et al., 2000; Yan, 2001; Sayar and Han, 2015 and 2016; Kendal et al., 2016; Erdemci, 2018).

The aims of the study were i-to determine and compare salt tolerance of the genotypes at germination and early seedling stages, ii-to determine the varieties that could be grown in the rice cultivation areas with the salinity problem and to recommend the varieties with the highest tolerance to salinity, iii-and to find out rice genotypes which could be used in future rice breeding programs to improve salt stress tolerance in 17 Turkish rice genotypes widely grown in Turkey.

MATERIALS AND METHODS

The 17 Turkish rice cultivars that were provided by Thrace Agricultural Research Institute of Turkey were used as plant materials. The genotypes used in the study were; Osmancik-97, Gonen, Halilbey, Edirne, Gala, Cakmak, Hamzadere, Efe, Pasali, Kale, Yatkin, Biga Incisi, Tosya Gunesi, Surek M711, Balaban, Sarhan and Ulfet. The seeds of the rice genotypes used in the experiment were surface sterilized with 10% sodium hypochlorite solution for five minutes and then rinsed with distilled water. The seeds were placed into 11 cm diameter petri dishes each having 25 seeds on a double-layer filter paper. Afterwards, 15 ml of relevant salt solution, including 0, 40, 80 and 120 mM NaCl were added to petri dishes for each salt concentration and put into the incubator at 25+-1 AdegC for eight days.

The investigated traits such as water uptake (WU-%), germination ratio (GR-%), root length (RL-cm), shoot length (SL-cm), root and shoot fresh weight (RFW and SFW-mg) and root and shoot dry weights (RDW and SDW-mg) were measured as described in the literature (Akbarimoghaddam et al., 2011). The plant samples were prepared to determine ion contents by burning at 500 AdegC for 6 h and the ashes were dissolved with 2 ml of 10 N Nitric Acid (HNO3). The readings were performed using Flame photometer (BWB Technologies) (Muftuoglu et al., 2014). The photosynthetic pigments were determined in fresh leaf samples before harvest. The fresh leaf samples (500 mg) were homogenized with acetone (90% v v-1), the homogenate was filtered and made up to a final volume of 10 ml. The absorbance of homogenate was measured at 663, 645, and 470 nm using a spectrometer (Shimadzu UV-1201; Tokyo).

Finally, the concentrations of chlorophyll a (Chl a), chlorophyll b (Chl b), chlorophyll a + b (Chl a + b), and carotenoids (Car) were calculated according to the formulas showed below by Lichtenthaler (1987).

Chl a (mg/g FW) = (11.75 x A663) - (2.35 x A645) x V/FW

Chl b (mg/g FW) = (18.61 x A645) - (3.96 x A663) x V/FW

Car (mg/g FW) = (1000 x A470) - (2.27 x Chl a) - (81.4 x Chl b)/227 x V/FW

Where; A: absorbance at wavelength, V: the final volume of acetone solution, and FW: the fresh weight of leaf samples in mg.

Statistical analyses: The experiment was arranged in a completely randomized factorial design with three replications. The experimental data was analyzed using ANOVA procedure with the JMP software (JMP, 2007). Duncan's Multiple Range Test was used to compare mean data. GGE biplot analysis was carried out using GenStat 14th edition as described by Yan (2001).

Table 1. Water uptake, germination rate, root lenght and shoot lenght of rice cultivars under different salinity levels.

###Water uptake (%)###Germination rate (%)###Root length (cm)###Shoot length (cm)

###Salinity levels (mM)

Cultivars###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean

Halilbey###31.93ab###28.96c-h###28.83c-j###28.70c-j###29.60a###76.0h-m###66.6l-p###24.0u###9.3v###44.0f###6.43g-l###3.50q###2.33r-t###1.16u-x###3.35f###4.16i-k###2.20p-s###1.30u-y###1.00w-y###2.16k

Osmancik-###30.30abc###29.63a-f###28.03c-n###27.70c-o###28.91ab###94.6a-d###96.0a-c###88.0a-h###18.6uv###74.3d###6.30g-m###4.83p###2.56q-s###0.90x###3.65ef###4.53e-j###4.43g-j###2.63o-q###1.03w-y###3.15d-g

97

Cakmak###29.66a-e###27.10e-r###26.20h-u###25.80k-v###27.19cd###100.0a###93.3a-e###74.6i-m###25.3u###73.3d###6.50f-l###6.60f-k###2.00s-w###0.70x###3.95cde###4.56e-j###4.36g-j###2.30p-s###0.96xy###3.05e-h

Gonen###26.50g-s###26.03i-v###25.10o-v###24.66p-v###25.57e###100.0a###93.3a-e###86.6b-i###53.3q-s###83.3bc###6.86d-i###8.36a-c###2.86q-s###0.93wx###4.75a###5.96a###5.40a-d###3.26m-o###1.03w-y###3.91a

Hamzadere###30.20a-d###29.63a-f###26.73g-s###23.60t-w###27.54bc###96.0a-c###96.0a-c###90.6a-f###48.0r-t###82.6c###6.13g-n###6.43g-l###2.13s-u###0.93wx###3.90de###5.96a###5.16b-e###2.53p-r###0.86xy###3.63ab

Efe###27.03e-s###25.96j-v###25.33m-v###25.10o-v###25.85de###97.3a-c###92.0a-f###90.6a-f###72.0k-o###88.0abc###5.06n-p###7.20d-g###3.26q-r###0.90x###4.10b-e###4.23i-k###5.03b-g###2.86n-p###0.86xy###3.25c-f

Gala###28.86c-i###28.36c-l###27.43c-p###26.43h-t###27.77bc###92.0a-f###81.3e-k###86.6b-i###41.3st###75.3d###9.2a###6.06h-n###2.56q-s###0.93wx###4.69a###5.46a-c###5.33a-d###2.60o-r###0.76y###3.54bc

Paali###30.10a-d###27.46c-p###27.00e-s###26.06i-v###27.65bc###96.0a-c###94.6a-d###85.3e-j###58.6p-r###83.6bc###4.93op###5.83i-p###3.30q-r###0.93wx###3.75ef###4.43g-j###4.30h-k###1.66s-w###0.80xy###2.80hi

Edirne###28.96c-h###28.90c-i###27.33d-q###26.36h-u###27.89bc###81.3e-k###61.3n-q###60.0o-r###26.6u###57.3e###8.83ab###5.50l-p###2.66q-s###1.03v-x###4.50ab###5.50ab###3.66k-m###1.93r-v###1.00w-y###3.02e-h

Balaban###26.90e-s###26.40h-t###24.90o-v###24.20s-v###25.60e###90.6a-f###89.3a-g###76.0h-m###74.6i-m###82.6c###6.86d-i###6.40g-l###2.56q-s###2.06s-v###4.47abc###3.40l-n###3.43l-n###1.36t-y###1.13w-y###2.33jk

Ulfet###30.10a-d###28.20c-m###27.36d-p###25.70k-v###27.84bc###82.6d-k###77.3g-l###64.0m-q###58.6p-r###70.6d###5.23m-p###5.70j-p###2.53q-s###2.16s-u###3.90de###4.23i-k###3.46l-n###1.46t-x###1.26v-y###2.60ij

Sarhan###26.16h-v###25.30n-v###24.86o-v###24.46q-v###25.20e###96.0a-c###92.0a-f###82.6d-k###82.6d-k###88.3abc###7.03d-h###6.73e-j###2.10s-v###2.36r-t###4.55ab###4.63e-j###4.06j-l###1.43t-y###1.40t-y###2.88ghi

Yatkin###28.53c-k###27.60c-o###27.13e-r###26.30h-u###27.39c###98.6ab###92.0a-f###88.0a-h###77.3g-l###89.0ab###5.63k-p###7.70c-e###2.86q-s###3.03q-s###4.80a###5.13b-f###4.93b-h###2.33p-s###2.00q-t###3.60ab

Biga ncisi###27.36d-p###26.76f-s###24.33r-v###23.30vw###25.44e###97.3a-c###96.0a-c###92.0a-f###77.3g-l###90.6a###6.76e-j###7.93b-d###3.30q-r###1.36t-x###4.84a###5.43a-d###5.46a-c###1.96q-u###1.10w-y###3.49bcd

Tosya###32.26a###27.70c-o###21.23w###20.93w###25.53e###81.3e-k###76.0h-m###73.3j-n###57.3p-r###72.0d###7.80b-e###7.53c-f###2.90q-s###1.03v-x###4.81a###5.03b-g###4.56e-j###2.70op###1.03w-y###3.33b-e

Gunei

Surek M711###26.26h-u###25.33m-v###24.90o-v###23.50uvw###25.00e###89.3a-g###88.0a-h###73.3j-n###40.0t###72.6d###6.56f-e###7.53c-f###2.16s-u###1.10u-x###4.34a-d###4.76d-i###4.80c-i###1.30u-y###0.80xy###2.91f-i

Kale###30.03a-d###29.36b-g###26.83e-s###25.53l-v###27.94bc###98.6ab###88.0a-h###80.0f-k###72.0k-o###84.6abc###6.73e-j###5.96h-o###2.10s-v###0.83x###3.90de###4.56e-j###4.46f-j###1.83s-v###0.76y###2.90f-i

Mean###28.89 a###27.57 b###26.09 c###25.19 d###26.93###92.2 a###86.6 b###77.4 c###52.5 d###77.1###6.64 a###6.46 a###2.60 b###1.31 c###4.25###4.82 a###4.41 b###2.08 c###1.04 d###3.08

LSD###SL:###CxSL:###SL:###CxSL:###SL:###CxSL:

###C:1.42###SL: 0.69###CxSL:2.84###C:6.08###C:0.53###C: 0.34

(P<0.05)###2.95###12.16###0.26###1.06###0.16###0.68

###CxSL:###CxSL:

F values###C: **###SL: **###CxSL: *###C:**###SL: **###**###C: **###SL: **###**###C:**###SL:**###CxSL:**

Table 2. Root and shoot fresh - dry weights of rice cultivars under different salinity levels.

###Root (fresh weight)###Root (dry weight)###Shoot (fresh weight)###Shoot (dry weight)

###Salinity levels (mM)

Cultivars###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean

Halilbey###16.14i-m###5.84r-y###3.81t-y###2.72v-y###7.13l###2.47h-n###1.42r-v###0.26zh###0.14zi###1.07g###11.42k-q###7.97t-w###4.43zd###2.39zg###6.55j###1.81l-r###1.67n-s###0.94xyz###0.44ze###1.22h

Osmancik-###17.49g-k###21.44e-h###8.14q-u###3.79t-y###12.71d-g###2.92d-j###2.45i-n###1.82o-s###0.62x-zc###1.95bcd###11.03n-r###13.22g-l###7.37v-x###3.80ze###8.85efg###2.10h-l###2.08h-m###1.57p-t###0.48zd###1.56g

97

Cakmak###17.84g-k###14.66j-o###5.69r-y###3.29u-y###10.37g-j###2.26l-p###1.81o-s###1.14t-x###0.64x-zc###1.46ef###13.60f-j###12.59h-n###8.53s-v###4.20zd###9.73de###2.17f-l###1.87k-q###1.50q-t###0.60zc###1.54g

Gonen###36.18a###24.11c-f###9.82o-s###1.78xy###17.97ab###3.94a###3.00d-i###1.65q-t###0.26zh###2.21ab###18.40ab###17.70b-d###11.07m-q###4.73zc###12.97a###3.24a###3.13a###2.02h-o###1.00v-y###2.35a

Hamzadere###18.68g-k###17.63g-k###5.18s-y###2.23w-y###10.93f-j###3.77ab###2.94d-i###1.29r-w###0.39zf###2.10abc###16.21c-e###14.07f-h###7.51u-x###4.10zd###10.47cd###2.91a-d###2.55c-f###1.79l-r###0.90xyz###2.04bc

Efe###15.27j-n###18.06g-k###11.90l-q###1.61xy###11.71e-i###2.87d-k###2.57g-n###2.05n-q###0.35zg###1.96bcd###12.16h-o###13.25g-l###9.49q-u###4.49zd###9.85de###2.24e-k###2.16f-l###1.90j-p###0.98w-z###1.82def

Gala###15.86i-m###22.65c-g###9.30p-s###1.55xy###12.34d-h###3.34b-e###3.00d-i###1.84o-r###0.26zh###2.11abc###11.92i-p###15.65d-f###8.64s-v###3.47ze###9.92cd###2.33e-i###2.24e-k###1.80l-r###0.93xyz###1.83de

Paali###19.89f-j###22.71c-g###10.49n-r###3.51u-y###14.15cde###3.09d-g###2.68f-m###1.60q-t###0.66x-zc###2.01bc###13.18g-m###13.72f-j###7.05v-y###4.48zd###9.61de###2.14g-l###2.11h-l###1.39s-v###0.93xyz###1.64efg

Edirne###15.07j-o###8.96p-t###2.82v-y###2.81v-y###7.42kl###2.52h-n###1.47r-u###0.46yze###0.39zf###1.21fg###13.99f-i###9.86p-t###5.53x-za###3.51ze###8.22gh###2.52d-g###2.11h-l###1.23t-x###0.36zf###1.55g

Balaban###11.39m-q###11.36m-q###5.88r-y###5.66r-y###8.57jkl###2.84d-k###2.11m-q###0.96u-z###0.99u-y###1.72de###8.93r-v###8.58s-v###5.02yzb###4.79zc###6.83ij###2.21f-k###1.97i-p###1.09u-y###0.87x-za 1.54g

Ulfet###17.04h-l###14.10k-p###5.54r-y###2.55w-y###9.81h-k###2.49h-n###2.36j-o###0.95u-z###0.59x-zd###1.60e###11.83j-p###9.52q-u###5.94w-z###3.45ze###7.69hi###2.04h-n###1.97i-o###1.37s-w###1.19t-x###1.65efg

Sarhan###19.38f-j###13.95k-p###2.45w-y###2.31w-y###9.52i-l###3.03d-h###2.23m-p###0.29zh###0.80w-zb###1.59e###13.86f-j###11.16l-q###4.76zc###4.32zd###8.53fgh###2.19f-l###2.27e-j###1.02v-y###1.01v-y###1.63fg

Yatkin###33.45a###34.86a###5.89r-y###7.18q-w###20.34a###3.78ab###2.96d-i###0.89v-za###1.59q-t###2.30a###15.28e-g###17.18b-e###7.76u-w###6.06 w-z###11.57b###2.39e-h###2.33e-i###1.45r-u###1.70m-s###1.96cd

Biga ncisi###31.53ab###27.21bc###5.00s-y###2.17w-y###16.48bg###3.69abc###2.86d-k###0.74w-zb###0.26zh###1.88cd###20.43a###18.26bc###9.58q-w###4.99yzb###13.31a###3.27a###2.98ab###1.64o-s###0.94xyz###2.21ab

Tosya###26.70b-d###21.89d-h###6.72q-x###3.22u-y###14.63cd###3.16c-f###2.82e-l###1.27s-w###0.49yze###1.93bcd###18.00bc###14.03f-i###7.48u-w###4.07zd###10.89bc###2.92abc###2.62b-e###1.43r-u###0.76yzb###1.93cd

Gunei

Surek###26.29b-e###15.07j-o###5.02s-y###1.01y###11.85e-i###3.40a-d###1.74p-s###0.81w-zb###0.03zj###1.49e###13.90f-j###10.26o-s###4.98yzb###2.83zf###7.99gh###2.05h-n###1.86k-q###0.76yzb###0.34zf###1.25h

M711

Kale###20.81f-i###21.84d-h###7.91q-v###3.34u-y###13.47def###2.54g-n###2.33k-o###1.15t-x###0.37zg###1.60e###13.43g-k###11.17l-q###8.78s-v###4.73zc###9.53def###2.04h-n###2.11h-l###1.45r-u###0.87x-za 1.61g

Mean###21.12 a###18.61 b###6.56 c###2.98 d###12.31###3.07 a###2.40 b###1.13 c###0.52 d###1.78###13.97 a###12.83 b###7.29 c###4.14 d###9.55###2.39 a###2.24 b###1.43 c###0.84 d###1.72

LSD###CxSL:###CxSL:###SL:###CxSL:

###C:2.57###SL: 1.24###CxSL:5.15###C:0.27###SL: 0.13###C:1.00###SL: 0.48###C: 1.98

(P<0.05)###0.55###2.02###0.10###0.39

###CxSL:###CxSL:

F values###C: **###SL: **###CxSL: **###C:**###SL: **###**###C: **###SL: **###**###C:**###SL:**###CxSL:**

Table 3. Shoot Na, K, Ca and K/Na of rice cultivars under different salinity levels.

###Na###K###Ca###K/Na

###Salinity levels (mM)

Cultivars###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean

Halilbey###1.39y###46.90mno###58.36gh###78.29d###46.24b###19.19j-n###17.27n-r###11.39zg###6.68zp###13.63h###1.43s###2.39k-o###3.62c-f###0.83t###2.07cd###13.77b###0.37l-r###0.19n-r###0.08qr###3.60b

Osmancik-###2.69y###32.62vwx###46.46m-p###136.32a###54.52a###23.73abc###25.55a###16.83o-s###6.87zo###18.24a###0.31u###2.07n-q###2.89hij###0.31u###1.39f###8.78i###0.78l###0.36l-r###0.05r###2.49hi

97

Cakmak###2.03y###35.39t-w###50.63j-m###69.76ef###39.45e###18.22m-p###22.59c-f###14.28t-x###6.53zr###15.40fg###0.23u###1.81qrs###2.13l-q###4.61b###2.20c###8.95i###0.64l-p###0.28l-r###0.09qr###2.49hi

Gonen###1.48y###33.57u-x###41.72pqr###48.88k-n###31.41i###15.75q-u###19.14j-n###17.12o-r###14.36t-x###16.59de###0.18u###1.79qrs###2.11m-q###2.91hij###1.75e###10.60ef###0.57l-q###0.41l-r###0.29l-r###2.97def

Hamzadere###1.66y###33.11vwx###48.70k-n###52.18i-l###33.91gh###19.71i-m###23.34bcd###16.69o-s###9.48zj###17.31a-d###0.18u###1.78qrs###2.55j-m###3.12ghi###1.91de###11.89cd###0.70lmn###0.34l-r###0.18o-r###3.28c

Efe###2.03y###30.83vwx###40.73qrs###74.35de###36.99f###20.61f-k###22.71cde###18.06m-p###11.50zf###18.22a###0.22u###1.94o-r###2.36k-p###3.74cde###2.07cd###10.14fg###0.73lm###0.44l-r###0.15pqr###2.87efg

Gala###1.82y###30.54wx###44.87n-q###84.07c###40.32de###17.45n-q###22.21c-h###15.98q-t###13.10w-zb###17.19bcd###0.03u###1.79qrs###2.59jkl###4.76ab###2.29bc###9.58h###0.73lm###0.35l-r###0.15pqr###2.70gh

Paali###1.83y###38.49r-u###61.81g###72.81ef###43.74c###19.60i-m###22.34c-g###13.37w-za###10.57zh###16.47de###0.05u###2.09m-q###3.09ghi###3.83cd###2.27bc###10.67e###0.58l-q###0.22n-r###0.14pqr###2.90efg

Edirne###2.16y###47.66l-o###63.00g###73.66def###46.62b###24.84ab###19.88i-m###15.41yzd###12.25###18.10ab###0.21u###2.52j-n###3.34e-h###3.89c###2.49b###11.56d###0.42l-r###0.24m-r###0.16pqr###3.10cde

Balaban###1.84y###38.42r-u###87.61c###84.29c###53.04a###14.44t-w###16.91o-s###8.81zk###8.63zl###12.19i###0.03u###2.17k-q###4.37b###5.15a###2.93a###7.82j###0.44l-r###0.10qr###0.10qr###2.11j

Ulfet###3.17y###39.85rst###69.08f###105.79b###54.47a###20.74e-j###21.56d-i###13.59v-z###12.44x-zc###17.08cd###0.27u###2.48j-n###3.54c-g###5.18a###2.87 a

###6.55k###0.54l-r###0.19n-r###0.12qr###1.85k

Sarhan###1.92y###31.70vwx###62.13g###73.06ef###42.20cd###18.52l-o###21.41d-i###16.03q-t###15.54q-v###17.88abc###0.15u###1.92pqr###3.20f-i###3.62c-f###2.22c###9.65gh###0.68l-o###0.26m-r###0.21n-r###2.70gh

Yatkin###1.97y###32.71vwx###59.05gh###71.00ef###41.18de###15.96q-t###18.64k-o###13.95u-y###13.30w-zb###15.47fg###0.21u###1.73qrs###3.15ghi###3.72cde###2.20c###8.08j###0.57l-q###0.24m-r###0.18o-r###2.27ij

Biga ncisi###1.09y###30.21x###47.15mno###49.71k-n###32.04hi###17.08o-r###22.10c-h###13.15w-zb###10.21zi###15.64efg###0.12u###1.54rs###2.52j-n###2.87ij###1.76 e

###15.7a###0.73lm###0.28l-r###0.20n-r###4.22a

Tosya###1.60y###35.77s-v###49.62k-n###54.82hij###35.45fg###16.49p-s###20.57g-k###14.94s-w###7.69zn###14.93g###0.14u###1.77qrs###2.59jkl###3.19f-i###1.92de###10.32ef###0.58l-q###0.30l-r###0.14pqr###2.83fg

Gunei

Surek###1.70y###33.27vwx###45.45n-q###55.82hi###34.06gh###20.62f-k###22.67cde###11.92ze###10.07zi###16.32def###0.03u###1.81qrs###2.61jk###3.39d-g###1.96de###12.09c###0.68l-o###0.26m-r###0.18o-r###3.30c

Table 4. Root Na, K, Ca and K/Na of rice cultivars under different salinity levels.

###Na###K###Ca###K/Na

###Salinity levels (mM)

Cultivars###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean

Halilbey###6.40zf###53.75tuv###96.56ij###132.05d###72.19c###22.13ab###9.77n-t###7.05v-z###5.05zc###11.00de###0.65v###3.78mn###7.09hi###5.59j###4.27d###3.48a###0.18h-m###0.07j-m###0.04lm###0.94ab

Osmancik-

97###6.24zf###24.43ze###37.21za###70.68opq###34.64gh###15.89c-g###11.40k-n###10.58l-q###6.16yz###11.01de###0.57v###2.04stu###2.78o-r###4.22lm###2.40j###2.56de###0.46g###0.29g-m###0.09j-m###0.85bcd

Cakmak###6.18zf###29.85zd###52.62uvw###64.02qrs###38.17fg###17.45c###11.99klm###12.19kl###5.78za###11.85bc###0.57v###2.43q-u###3.40no###3.72mn###2.53j###2.97b###0.40ghi###0.23g-m###0.09j-m###0.92abc

Gonen###4.45zf###22.81ze###38.12z###70.66pq###34.02h###8.86r-u###7.61u-y###8.94q-u###8.83r-u###8.56g###0.41v###1.87u###2.58q-t###4.67kl###2.38j###1.99f###0.33g-j###0.23g-m###0.12j-m###0.67efg

Hamzadere###4.51zf###56.17s-v###55.32tuv###110.51ef###56.63d###11.65klm###10.54l-q###15.36e-i###9.15p-u###11.68bcd###0.43v###2.51q-u###4.68kl###3.00opq###2.65ij###2.58de###0.19h-m###0.28g-m###0.08j-m###0.78de

Efe###5.62zf###38.70yz###65.17qr###107.01efg###54.13d###16.22c-f###11.64klm###11.73klm###6.86w-z###11.62bcd###0.55v###2.79o-r###3.72mn###7.21ghi###3.57f###2.89bc###0.31g-l###0.18h-m###0.06klm###0.86bcd

Gala###4.81zf###61.47rst###41.88yz###85.35lmn###48.38e###15.71d-h###11.37k-n###12.19kl###3.04ze###10.57e###0.45v###2.84o-r###3.83mn###6.77i###3.47f###3.32a###0.18h-m###0.29g-m###0.04lm###0.96ab

Paali###6.08zf###42.82xy###45.43wxy###132.04d###56.60d###16.86cde###10.35m-r###10.64l-p###9.91n-s###11.94bc###0.58v###2.65p-s###3.79mn###8.58bcd###3.90e###2.77bcd###0.24g-m###0.24g-m###0.08j-m###0.83bcd

Edirne###7.93zf###60.39r-u###106.50e-h###113.85e###72.17c###22.77a###13.90ij###5.47za###6.38yz###12.13ab###0.73v###4.17lm###7.21ghi###8.49cd###5.15c###2.87bc###0.23g-m###0.05lm###0.06klm###0.80cde

Balaban###5.62zf###91.43jkl###92.26jkl###99.39g-j###72.18c###10.61l-p###11.06k-o###8.24t-w###7.74u-x###9.41f###0.36v###3.88mn###7.79efg###9.14bc###5.29bc###1.88f###0.12j-m###0.09j-m###0.08j-m###0.54g

Ulfet###8.70zf###94.15jk###103.04f-i###160.12b###91.51a###23.29a###14.08hij###8.89r-u###5.06zc###12.83a###0.78v###4.10lm###8.06def###11.78a###6.18a###2.67cde###0.15h-m###0.09j-m###0.03m###0.73def

Sarhan###7.08zf###86.71klm###141.96c###97.10ij###83.21b###17.34cd###12.52jk###8.41s-w###8.61s-v###11.71bcd###0.62v###3.84mn###9.19a###8.44de###5.52b###2.45e###0.14i-m###0.06klm###0.09j-m###0.68ef

Yatkin###6.98zf###52.36uvw###78.42nop###208.66a###86.61b###14.75f-i###9.56o-t###10.68l-p###9.74n-t###11.18cde###0.57v###2.34r-u###5.71j###7.63fgh###4.06de###2.11f###0.18h-m###0.14i-m###0.05lm###0.62fg

Biga ncisi###4.13zf###22.94ze###61.02rst###78.90mno###41.75f###14.55ghi###10.98k-o###6.46yz###3.06ze###8.76fg###0.42v###1.96tu###3.87mn###5.57j###2.95hi###3.52a###0.48g###0.10j-m###0.04lm###1.04a

Tosya

Gunei###7.59zf###50.37vwx###67.93qr###91.43jkl###54.33d###15.65e-h###9.20p-u###6.78xyz###5.98z###9.40f###0.60v###2.10stu###3.28nop###6.57i###3.14gh###2.07f###0.18h-m###0.10j-m###0.07j-m###0.60fg

Surek

M711###6.01zf###34.47zb###66.85qr###104.67f-i###52.99d###16.84cde###11.31k-n###9.50o-t###5.13zb###10.69e###0.53v###2.39q-u###4.10lm###5.07jk###3.02gh###2.81bcd###0.32g-k###0.14i-m###0.05lm###0.83bcd

Kale###5.94zf###29.90zc###55.60tuv###98.60hij###47.51e###20.74b###12.52jk###6.56yz###4.13zd###10.98de###0.51v###2.41q-u###3.69mn###6.67i###3.32fg###3.49a###0.42gh###0.12j-m###0.04lm###1.02a

Mean###6.13 d###50.16 c###70.93 b###107.36 a###58.64###16.55 a###11.17 b###9.39 c###6.51 d###10.90###0.55 d###2.83 c###4.99 b###6.65 a###3,75###2.73 a###0.27 b###0.16 c###0.06 d###0.80

LSD###CxSL:###C:###SL:###C:###SL:###CxSL:

###C: 4.14###SL: 2.01###CxSL:8.29###C:0.83###SL: 0.40###CxSL:0.66

(P<0.05)###1.66###0.32###0.16###0.14###0.07###0.27

###CxSL:

F values###C: **###SL: **###CxSL: **###C:**###SL: **###**###C: **###SL: **###CxSL: ** C:**###SL:**###CxSL:**

Table 5. Chl a, Chl b, Chl a + b and Carotenoid of rice cultivars under different salinity levels.

###Chla###Chlb###Chla+b###Caratenoid

###Salinity levels (mM)

Cultivars###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean###0###40###80###120###Mean

Halilbey###0.210zn###0.254zh###0.140zac###0.190zr###0.199q###0.069za###0.114mn###0.046zg###0.214c###0.111e###0.280zm###0.368zb###0.186zv###0.404x###0.310o###0.137zc###0.157y###0.094zk###0.111zf###0.124o

Osmancik-

97###0.246zi###0.414g###0.215zm###0.131zae###0.251o###0.089uv###0.108p###0.079x###0.109p###0.096j###0.336zf###0.522hi###0.294zj###0.240zr###0.348l###0.154z###0.243f###0.130zd###0.240zr###0.147n

Cakmak###0.342s###0.372l###0.222zl###0.102zai###0.260m###0.097r###0.092st###0.068zb###0.063zd###0.080l###0.440u###0.464p###0.290zl###0.165zac###0.340m###0.211m###0.226ij###0.137zc###0.065zp###0.160k

Gonen###0.265ze###0.552a###0.369m###0.125zaf###0.328c###0.067zc###0.115m###0.096r###0.043zh###0.080l###0.332zg###0.667b###0.465p###0.168zab###0.408e###0.171vw###0.327a###0.225j###0.085zm###0.202 b

Hamzadere###0.264zf###0.504b###0.368mn###0.193zp###0.332b###0.073z###0.142f###0.097r###0.121l###0.108f###0.337zf###0.646c###0.465p###0.314zh###0.441c###0.170w###0.301b###0.222k###0.117ze###0.202b

Efe###0.292z###0.481c###0.335t###0.164zt###0.318d###0.112no###0.127j###0.123kl###0.089uv###0.113d###0.405x###0.608d###0.458q###0.253zo###0.431 d###0.191s###0.284c###0.207n###0.104zh###0.196 c

Gala###0.306x###0.390i###0.265ze###0.272zc###0.308f###0.134hi###0.132i###0.092st###0.248b###0.151b###0.440u###0.522hi###0.358zc###0.520i###0.460b###0.194r###0.233h###0.163x###0.162x###0.188d

Paali###0.383k###0.448d###0.362q###0.180zs###0.344a###0.108p###0.124k###0.121l###0.112no###0.116c###0.492lm###0.572e###0.484n###0.293zk###0.460b###0.236g###0.266d###0.213m###0.108zg###0.205a

Edirne###0.342s###0.333u###0.283za###0.202zo###0.290i###0.111o###0.083w###0.064zd###0.207d###0.116c###0.453r###0.416v###0.347ze###0.410w###0.407ef###0.216l###0.204op###0.182u###0.128zd###0.182g

Balaban###0.366no###0.422e###0.262zg###0.145zab###0.299h###0.128j###0.104q###0.091tu###0.074z###0.099i###0.495kl###0.526g###0.354zd###0.219zt###0.398g###0.232h###0.253e###0.164x###0.095zk###0.186e

Ulfet###0.302y###0.275zb###0.277zb###0.155zu###0.253n###0.097r###0.093st###0.112no###0.105q###0.102h###0.400y###0.369zb###0.390z###0.260zn###0.355k###0.192rs###0.171vw###0.172v###0.096zj###0.157l

Sarhan###0.292z###0.393h###0.365op###0.156zu###0.301g###0.092st###0.105q###0.135gh###0.087v###0.105g###0.384za###0.498jk###0.500j###0.243zp###0.406f###0.184u###0.236g###0.223k###0.097zi###0.184f

Yatkin###0.237zk###0.388ij###0.324v###0.134zad###0.271k###0.072z###0.136g###0.128j###0.049zf###0.096j###0.309zi###0.524gh###0.452rs###0.183zy###0.367j###0.151za###0.228i###0.189t###0.088zl###0.163j

Biga ncisi###0.349r###0.417f###0.246zi###0.114zag###0.281j###0.152e###0.128j###0.068zb###0.037zi###0.096j###0.501j###0.545f###0.314zh###0.150zae###0.377h###0.217l###0.245f###0.155yz###0.078zn###0.173h

Tosya

Gunei###0.364pq###0.342s###0.217zm###0.149zy###0.268l###0.094s###0.108p###0.141f###0.076y###0.105g###0.458q###0.450st###0.358zc###0.225zs###0.373i###0.228i###0.203p###0.145zb###0.094zk###0.167i

Surek M711###0.344s###0.363pq###0.152zv###0.386j###0.311 e###0.135gh###0.128j###0.055ze###0.462a###0.195a###0.479o###0.491m###0.208zu###0.848a###0.506a###0.206no###0.216l###0.097zi###0.227ij###0.186e

Kale###0.312w###0.241zj###0.266zd###0.106zah###0.232 p###0.135gh###0.112no###0.078xy###0.050zf###0.094k###0.448t###0.353zd###0.345ze###0.156zad###0.325n###0.198q###0.182u###0.164x###0.071za###0.153m

Mean###0.307 b###0.388 a###0.275 c###0.171 d###0.285###0.104 c###0.115 b###0.094 d###0.126 a###0.110###0.411 b###0.502 a###0.369 c###0.297 d###0.395###0.193 b###0.234 a###0.169 c###0.105 d###0.175

LSD###C:###SL:###CxSL:###C:###SL:###CxSL:###C:###SL:###CxSL:###C: 0.001###SL:###CxSL:

(P<0.05)###0.0010###0.0005###0.0021###0.0010###0.0005###0.0021###0.0016###0.0007###0.0031###0.0005###0.002

###CxSL:###CxSL:

F values###C: **###SL: **###**###C:**###SL:**###CxSL:**###C: **###SL: **###**###C: **###SL: **###CxSL:**

RESULTS AND DISCUSSION

Germination and Seedling Traits: The water uptake of seeds was significantly affected by salinity levels and their interactions (Table 1). At the end of 24 h for mean values, the highest water uptake rate was 29.60% by Halilbey genotype, while the lowest water uptake rate was 25% by Surek M711 genotype. The average water uptake rate was 26.93% among the rice genotypes. Salt concentrations had a negative effect on water uptake rate at 24 h, and there was a decrease water uptake with an increase in salt concentrations. According to mean salinity levels; the highest water uptake rate (28.89%) was found in control treatment while the lowest water uptake (25.19%) was found for 120 mM salt concentration (Table 1). For germination rate, NaCl concentrations and genotype x salt concentration interactions varied between 9.3 and 100%. The highest germination rate was obtained from Cakmak and Gonen varieties with 100% germination rate at control level.

In contrast, the lowest germination rate (9.3%) was noted for Halilbey variety at 120 mM NaCl dose. Among the varieties, Biga Incisi (90.6%) was the variety that had lowest effect by salt stress, and Halilbey (44%) was the most sensitive variety. The decrease in germination rates of some varieties were declined gradually with increase in NaCl doses, the reduction rates in some varieties were more severe and irregular (Table 1). The rice cultivars were found variable for root length. The longest root lengths were obtained from Biga Incisi, Tosya Gunesi, Yatkin, Gonen and Gala cultivars as 4.84, 4.81, 4.80, 4.75 and 4.69 cm, respectively while the shortest root length was obtained from Halilbey cultivar as 3.35 cm. The root length was gradually decreased by increased salt concentration and the reduction was recorded as 80.27% at 120 mM salt concentration compared to control group. Genotype x salt concentration interaction was identified as significant for root length.

Therefore, Edirne cultivar had the highest root length at 0 mM salt concentration with 8.83 cm, while Cakmak cultivar had the lowest root length at 120 mM salt concentration with 0.70 cm (Table 1). According to shoot lenght analyses, the highest shoot length was obtained from Gonen cultivar as 3.91 cm while the shortest shoot length was obtained from Halilbey cultivar as 2.16 cm. The shoot length was decreased significantly as a result of increasing salt concentration doses, and the shoot length was decreased by 78.42% at 120 mM salt concentration compared to control dose. Genotype x salt concentration dose interaction was significant on shoot length, while Gonen and Hamzadere varieties had the highest shoot length with 5.96 cm at 0 mM salt concentration, Gala and Kale varieties had the lowest shoot length values of 0.76 cm at 120 mM salt concentration (Table 1).

The root fresh weight was found significant among the rice cultivars and the highest value was obtained from Yatkin cultivar (20.34 mg), while Halilbey had the lowest value with 7.12 mg. In addition, the root fresh weight was decreased with the increased salt concentrations and 120 mM dose had 85.89% decrease compared to control. The genotype x salt concentration interaction was found important for root fresh weight and Gonen cultivar had 36.18 mg root fresh weight at control dose, while Surek M711 cultivar had the lowest root fresh weight with 1.01 mg at 120 mM dose (Table 2). There were significant differences among the rice cultivars and genotype x salt concentration interaction for shoot fresh weight. The highest shoot fresh weight values were obtained from Gonen and Biga Incisi cultivars as 12.97 and 13.31 mg, respectively and the lowest value was found as 6.55 mg from Halilbey cultivar.

The shoot fresh weight value of the cultivars decreased by increased salt concentrations and according to salt concentrations, a decrease of 70.36% was measured between 120 mM to control dose. The Biga Incisi cultivar had the highest shoot fresh weight with 20.43 mg at control dose, while Halilbey cultivar had the lowest shoor fresht weight with 2.39 mg at 120 mM dose (Table.2). The rice cultivars and genotype x salt concentration interaction were differed for root dry weight (RDW) (p <0.01). The highest RDW was obtained from Yatkin cultivar with 2.3 mg, while Halilbey cultivar had the lowest RDW with 1.07 mg. There was an 83.06% decrease between 120 mM and control doses. The Gonen cultivar had the 3.94 mg RDW at 0 mM dose, while Surek M711 had the 0.03 mg at 120 mM dose (Table 2). The significant differences were identified between rice genotypes and genotype x salt concentration interactions for shoot dry weight (SDW).

While the Gonen cultivar had the highest SDW with 2.35 mg, and the Halilbey cultivar had the lowest SDW with 1.22 mg. The decrease between 120 mM to 0 mM salt concentrations was measured as 65%. The Biga Incisi cultivar had the highest SDW with 3.27 mg at 0 mM concentration, while Surek M711 cultivar had the lowest SDW with 0.34 mg at 120 mM dose (Table 2). The Na and Ca rates were increased with high salt concentration in both root and shoot, while K and K/Na concentrations decreased which was observed in all genotypes and lower Na and Ca levels were accumulated in the shoots rather than roots. This result might be related with the direct relationship between root and NaCl. The lowest and highest Na concentrations were measured in Gonen (34.02 mg/g) and Ulfet (91.51 mg/g) cultivars in the roots, respectively. While the highest Na was observed in Balaban, Ulfet and Osmancik-97 (53.04, 54.47 and 54.52 mg/g, respectively) cultivars, the lowest Na was measured at Gonen (31.41 mg/g) in shoots.

On the other hand, the highest K value was observed in Ulfet cultivar with 12.83 mg/g and the lowest K concentration was obtained from Gonen with 8.56 mg/g in roots, while the highest K value was measured in Osmancik-97 cultivar with 18.24 mg/g. In addition, Balaban cultivar has the lowest K with 12.19 mg/g in shoots. In terms of Ca concentration in roots, Ulfet cultivar had the highest value with 6.18 mg/g and Gonen had the lowest value with 2.38 mg/g. While Balaban had the highest values as 2.93 mg/g, Osmancik-97 had the lowest Ca concentration as 1.39 mg/g in shoots. In addition, the highest K/Na value was obtained from Biga Incisi cultivar with 1.04 and the lowest one was obtained from Balaban cultivar with 0.54 in roots. Biga Incisi cultivar had the highest K/Na concentration with 4.22, while Ulfet cultivar had the lowest one with 1.85 in shoots (Table 3 and 4).

According to these results, osmotic pressure was increased with an increasing in salt concentrations, which decreased the water uptake of the seeds. Salinity inhibits water uptake of the seeds and this has been reported by Akbarimoghaddam et al. (2011) for wheat, Shereen et al. (2011) and Balkan et al. (2015) for rice and, Dogan and Carpici (2015; 2016) for wheat and triticale. Ekmekci et al. (2005) reported that reduction in germination rates because of increased salt levels was due to toxicity of Na + and Cl-ions. Furthermore, water imbibition required for the seed germination was blocked due to increased osmotic potential. Our results were in agreement with the findings of other researchers who have reported that the germination rates of genotypes were decreased with increasing salt concentrations in major crops (Dumlupinar et al., 2007; Datta et al., 2009; Akbarimoghaddam et al., 2011; Hussain et al., 2013; Mahmoodzadeh et al., 2013; Balkan et al., 2015; Solangi et al., 2015 and Zafar et al., 2015).

For the root length, similar results were reported by Atis (2011) for sorghum, Uyanik et al. (2014) for canola, Balkan et al. (2015) for rice, Solangi et al. (2015) for rice and Zafar et al. (2015) for rice, Dolo et al. (2016) for rice. However, Hussain et al. (2013) indicated that root length in rice was increased by higher salt concentration which was inconsistent to our findings. As the salt concentrations were increased, the length of the shoots was decreased. This situation was a result of negative effects on water uptake caused by the toxic effects of the salt ions and the osmotic pressure. Similar results were reported for sorghum (Atis, 2011), canola (Uyanik et al., 2014), and rice (Hussain et al., 2013; Balkan et al., 2015; Solangi et al., 2015). The root fresh weight values were lowered by higher salt concentrations. Similar findings were reported by Balkan et al. (2015) and Solangi et al. (2015) in rice.

We found that shoot fresh weight values were decreased with increased salt concentrations. Our findings were in agreement with Haq et al. (2009) for rice, Atis (2011) for sorghum, Hussain et al. (2013) for rice, Uyanik et al. (2014) for canola, Balkan et al. (2015) for rice, Solangi et al. (2015) for rice and Zafar et al. (2015) for rice. The results indicated that root growth was significantly decreased under severe salinity stress. Our results were also in agreement with the findings of Kazemi and Eskandari (2011), Balkan et al. (2015), Solangi et al. (2015), Zafar et al. (2015) and Dolo et al. (2016) who found that RDW was inhibited by high salinity levels in rice. It is also determined that high salt concentrations lowered the shoot dry weight which was supported by the results of the previous works in rice (Haq et al., 2009; Balkan et al., 2015; Solangi et al., 2015; Zafar et al., 2015).

In previous works; Tatar and Gevrek (2007) reported that increased salt concentrations caused an increase in leaf K/Na rates. Haq et al. (2009) also indicated that increased salt concentrations raised the Na ion accumulation in the shoots, while decreased the K ion accumulation. It is concluded that, one of the key features of plant salt tolerance is the ability of plant cells to maintain optimal K/Na ratio in the cytosol, when exposed to salt stress (Maathuis and Amtmann, 1999; Carden et al., 2003; Tester and Davenport, 2003; Ashraf, 2004; Kaya et al., 2012).

Photosynthetic Pigment Contents: Genotype, salt concentrations and genotype x salt concentration interactions were found significant for photosynthetic pigments. The Chl a pigment value was increased at 0 and 40 mM doses, while decreased at 80 and 120 mM salt concentration doses. The highest value obtained from Halilbey with 0.199 mg/g, while the lowest one obtained from Pasali cultivar with 0.344 mg/g. On the other hand, the Chl b pigment content was increased at control and 40 mM doses, while decreased at 80 mM doses, but had the highest content 120 mM dose. The highest Chl b pigment content at 120 mM was due to closed leaves of the shoots. Surek M711 had the highest content with 0.195 mg/g, and the lowest ones were obtained from Cakmak and Gonen cultivars with 0.080 mg/g. In addition, caratenoid pigment contents increased at control and 40 mM doses and decreased at 80 and 120 mM salt concentration doses.

The highest carotenoid value was obtained from Pasali cultivar with 0.205 mg/g, while Halilbey cultivar had 0.124 mg/g as the lowest. The Chl a + b content was higher at control and 40 mM doses, in contrast, it was decreased by 80 and 120 mM doses. The Surek M711 and Haliley cultivars indicated the highest (0.506 mg/g), and the lowest (0.310 mg/g) values, respectively (Table 5). It was determined that Chl a was much more affected than Chl b under salt stress which was also indicated by Lutts et al. (1996) and Tatar et al. (2010). It was also reported that chlorophyll pigment concentration was significantly changed under different salinity concentrations (Tatar and Gevrek, 2007; Aliu et al., 2015).

Evaluation of Genotype and Traits using GGE Biplot Analysis under Salt Stress: The GGE biplot analysis was performed to present a visual demonstration of the study. In recent years, the GGE biplot analysis was preferred by plant breeders and it provides interpretation of releationships between genotypes and environments, investigated traits and environments and correlations of investigated traits (Kendal et al., 2016; Aktas et al., 2017; Erdemci, 2018). The biplot can be used reliably and provided the explaining an adequate amount ([greater than or equal to]50%) of total variation (Yan et al., 2000). Accordingly, the results of the present study revealed that principal component analysis explained 56.41% of the total variation where 38.02% was represented by PC1 and 18.39% by PC2, respectively (Figure 1, 2 and 3). The biplot analysis showed the most promising characters as a selection criteria for the cultivars used in this study.

As shown in Figure 1, the genotypes and investigated traits were separeted by 5 mega-environments and differed in the distribution of genotypes. The Gonen variety found on the corner of the polygon in Figure 1 (which-won-where) shows the relationship between the genotypes and the features examined, the genotype has the highest values for SFW, RFW, SL, SDW, RDW, GR, RL, Chl a, Chl a + b, Car and shoot K. Halilbey was located on the corner of the polygon, but has no ideal properties for any characters. The genotypes at the corner of the polygon were found to have the most ideal or undesirable properties in terms of their characters. The ranking model of GGE biplot showed genotypes based on mean and stability of traits in Figure 2. The comparison model of GGE biplot also showed ideal genotypes based on traits in Figure 3. According to ranking and comparison of genotypes based on their traits (Figure 2 and 3), Gonen was an ideal genotype.

Furthermore, these genotypes can also be considered as ideal since its values for almost investigated traits. Other researchers have reported similar results for wheat, triticale and chickpea (Kendal et al., 2016; Aktas et al., 2017; Aslan et al., 2017, Erdemci, 2018).

Conclusion: As a result of the study, the salt stress affected germination and seedling stages of the rice genotypes. The increased salt doses gradually decreased the seed germination and limited the rice germination and seedling development stages. The rice cultivars responsed differently to the salinity stress and the Gonen cultivar showed the best performance in response to salt stress for the investigated traits; thus, it might be considered to be a good parent in breeding programs and salt stress tolerance studies to be conducted under field or in vitro conditions. It is also concluded that Gonen cultivar could be recommended for the rice growing saline lands.

Acknowledgements: This study was supported by the Scientific Research Council of Duzce University (DU BAP Project No: 2016.11.04.454). Authors would like to acknowledge Dr. Khawar Jabran for proof reading of the manuscript.

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