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Molecular Soil Biology (online), 2011, Vol.3 No.1ISSN1925-2005 http://msb.sophiapublisher.comBold’s Basal Medium (BBM): NaNO 3 0.250 g, NaCl0.025 g, K 2 HPO 4 0.075 g, KH 2 PO 4 0.175 g, H 3 BO 30.011 g, EDTA 0.050 g, KOH 0.031 g, Co(NO 3 ) 2·6H 2 O0.490 mg, CuSO 4 1 mg, MnCl 2·4H 2 O 1.440 mg,ZnSO 4·7H 2 O 8.820 mg , MoO 3 0.710 mg, MgSO 4·7H 2 O0.075 g, CaCl 2·2H 2 O 0.025 g, FeSO 4·7H 2 O 5 mg,distilled water 1 L, and the pH of the medium adjustedto 8.0.3.2 Methods3.2.1 Identification and purification of the algae5 g of soil was weighed and placed in 150 mLErlenmeyer flask containing 50 mL sterile water, andthen incubated in the culture room, the light intensityis 4 000~6 000 lux, photoperiod is 14 h: 10 h, 24 ℃ .When the algae appeared, we picked a fluid with ainoculating loop, and then streaked on the BBM solidmedium until individual colonies grown on the plates,and single colonies were purified more than 8 times.Figure 2 Analysis of algae expressed in different NaHCO 3stressesNote: Algae species included JB1~JB22 (except JB12) andCK1~CK8, incubated as described in Materials and Methods.Serial dilutions were spotted onto solid BBM mediumsupplemented without or with additional NaHCO 3 (100 mmol/L,200 mmol/L and 400 mmol/L). Growth was observed one week later.The isolation and screening of special resistancerelatedalgae species as the foundation of resistancerelatedgenes have an important theoretical andpractical significance on soil salinity remediation,ecological restoration and the promotion ofresistance-related genetic engineering of plants.3 Materials and Methods3.1 MaterialsSoil samples were collected from alkali spot soil inMay and September, 2009, in Anda area, HeilongjiangProvince, In addition, we selected garden nursery soilin Northeast Forestry University as the control.3.2.2 Resistance analysis of algaeSingle colonies of algae species purified was pickedinto liquid BBM medium, when the algae solution wasdense, took some vigorous solution, centrifuged,abandoned the supernatant, then added a certainamount of sterile deionized water, make sure theinitial of the cell concentration was 100 mg/mL,recorded as 5 -0 , and then diluted to 5 -1 , 5 -2 , 5 -3 , 5 -4 inturn. 4 μL of serial dilutions (JB9, which is filamenttous,was only picked filaments on the solid mediumwith no concentration gradient) were spotted ontosolid BBM medium supplemented without or withadditional NaCl (BBM medium whose pH wasadjusted to 8.0 as the basic culture and the controlmedium) and NaHCO 3 (BBM medium whose pH wasnot adjusted as the basic culture and the controlmedium). Growth was observed one week later.Authors’ contributionsJW and WS designed and conducted this experiment; TT participated theexperiment design and data analysis; SKL is the person who takes charge ofthis project, including experiment design, data analysis, writing andmodifying of the manuscript. All authors have read and approved the finalmanuscript.AcknowledgementsThis work was supported by the Heilongjiang Provincial Program forDistinguished Young Scholars (JC200609) and State ForestryAdministration 948 Program of PR China (No. 2008429) to Shenkui Liu.Authors appreciate two anonymous reviewers for their useful critical- 3 -