生态环境学报
生態環境學報
생태배경학보
ECOLOGY AND ENVIRONMENT
2013年
12期
1936-1944
,共9页
靳振江%黄海涛%刘杰%林华%张学洪
靳振江%黃海濤%劉傑%林華%張學洪
근진강%황해도%류걸%림화%장학홍
重金属%人工湿地%微生物群落%硝化%反硝化
重金屬%人工濕地%微生物群落%硝化%反硝化
중금속%인공습지%미생물군락%초화%반초화
heavy metals%constructed wetland%microbial community%nitrifying%denitrifying
在铬、铜和镍复合污染的3段波形潜流人工湿地中采集0~15 cm土壤样品，分别以单氨加氧酶的编码基因amoA和亚硝酸还原酶的编码基因nirK的部分序列作为硝化细菌和反硝化细菌的指示基因，采用实时聚合酶链式反应(实时PCR)与PCR-变性梯度凝胶电泳(PCR-DGGE)结合克隆和测序的方法，研究了硝化细菌与反硝化细菌群落结构的空间分布。结果表明，在人工湿地中，amoA和nirK基因丰度均在第3段最高，分别为(8.60±2.75)×106拷贝·g-1和（137.76±42.16）×106拷贝·g-1。回收的15个amoA基因序列中有7个属于簇3a，而8个nirK基因序列中有3个属于簇III。相关性分析表明，amoA基因拷贝数与全量铜、全量镍和铬的活化率呈显著负相关(r分别为-0.870、-0.884和-0.898)，但有效铜与amoA香农-维纳指数和辛普森指数之间均呈显著正相关(r分别为0.892和0.940)，铜的活化率与amoA基因均匀度呈显著正相关(r=0.887)；nirK基因拷贝数与全量铜、全量镍和铬的活化率也均呈显著负相关(r分别为-0.818、-0.820和-0.860），但有效铜与nirK基因均匀度呈显著正相关(r=0.917)。以上结果表明，硝化细菌和反硝化细菌在人工湿地各个阶段的变化规律基本相同，这二者的丰度与多样性指数对重金属污染的反应总体上呈相反的变化。
在鉻、銅和鎳複閤汙染的3段波形潛流人工濕地中採集0~15 cm土壤樣品，分彆以單氨加氧酶的編碼基因amoA和亞硝痠還原酶的編碼基因nirK的部分序列作為硝化細菌和反硝化細菌的指示基因，採用實時聚閤酶鏈式反應(實時PCR)與PCR-變性梯度凝膠電泳(PCR-DGGE)結閤剋隆和測序的方法，研究瞭硝化細菌與反硝化細菌群落結構的空間分佈。結果錶明，在人工濕地中，amoA和nirK基因豐度均在第3段最高，分彆為(8.60±2.75)×106拷貝·g-1和（137.76±42.16）×106拷貝·g-1。迴收的15箇amoA基因序列中有7箇屬于簇3a，而8箇nirK基因序列中有3箇屬于簇III。相關性分析錶明，amoA基因拷貝數與全量銅、全量鎳和鉻的活化率呈顯著負相關(r分彆為-0.870、-0.884和-0.898)，但有效銅與amoA香農-維納指數和辛普森指數之間均呈顯著正相關(r分彆為0.892和0.940)，銅的活化率與amoA基因均勻度呈顯著正相關(r=0.887)；nirK基因拷貝數與全量銅、全量鎳和鉻的活化率也均呈顯著負相關(r分彆為-0.818、-0.820和-0.860），但有效銅與nirK基因均勻度呈顯著正相關(r=0.917)。以上結果錶明，硝化細菌和反硝化細菌在人工濕地各箇階段的變化規律基本相同，這二者的豐度與多樣性指數對重金屬汙染的反應總體上呈相反的變化。
재락、동화얼복합오염적3단파형잠류인공습지중채집0~15 cm토양양품，분별이단안가양매적편마기인amoA화아초산환원매적편마기인nirK적부분서렬작위초화세균화반초화세균적지시기인，채용실시취합매련식반응(실시PCR)여PCR-변성제도응효전영(PCR-DGGE)결합극륭화측서적방법，연구료초화세균여반초화세균군락결구적공간분포。결과표명，재인공습지중，amoA화nirK기인봉도균재제3단최고，분별위(8.60±2.75)×106고패·g-1화（137.76±42.16）×106고패·g-1。회수적15개amoA기인서렬중유7개속우족3a，이8개nirK기인서렬중유3개속우족III。상관성분석표명，amoA기인고패수여전량동、전량얼화락적활화솔정현저부상관(r분별위-0.870、-0.884화-0.898)，단유효동여amoA향농-유납지수화신보삼지수지간균정현저정상관(r분별위0.892화0.940)，동적활화솔여amoA기인균균도정현저정상관(r=0.887)；nirK기인고패수여전량동、전량얼화락적활화솔야균정현저부상관(r분별위-0.818、-0.820화-0.860），단유효동여nirK기인균균도정현저정상관(r=0.917)。이상결과표명，초화세균화반초화세균재인공습지각개계단적변화규률기본상동，저이자적봉도여다양성지수대중금속오염적반응총체상정상반적변화。
The topsoil samples (0-15 cm) were collected in a three-stage wavy subsurface constructed wetland (W-SFCW) contaminated with heavy metals (Cr, Cu and Ni). The partial encoding genes of ammonia monooxygenase (amoA) and nitrite reductase (nirK) were used as the indicators of nitrifying and denitrifying bacteria, respectively. Spatial distributions of community abundance, composition and diversity of both bacteria were investigated through real-time polymerase chain reaction (real-time PCR) and PCR-denaturing gradinent electrophoresis (PCR-DGGE) combining cloning and sequencing. The abundance of amoA ranged from(1.32±0.43)×106 copies·g-1 to(8.60±2.75)×106 copies·g-1and nirK ranged from(5.78±1.15)×106 copies·g-1 to (137.75±42.16)×106 copies·g-1. The 7 of 15 amoA bands retrieved from DGGE gels were grouped in cluster 3a and 3 of 8 nirK bands were grouped in cluster III. A negative correlation between the total content of Cu and Ni and the activation rate of Cr and the amoA abundance (r=-0.870,-0.884 and-0.898),but a positive correlation between the Shannon-Weaver indices and Simpson indices of amoA and the available Cu (r=0.892 and 0.940), as well as the Evenness of amoA and the activation rate of Cu (r=0.887). A negative correlation between the total content of Cu and Ni and the activation rate of Cr and nirK abundance (r=-0.818,-0.820 and-0.860) was also detected, but a positive correlation between the nirK Evenness and the available Cu (r= 0.917). The above results indicated that nitrifying bacteria community responsed similar with the denitrifying bacteria community and microbial abundances changed generally contrary to diversities in wetland contanminated with heavy metals.
