海洋科学
海洋科學
해양과학
Marine Sciences
2015年
9期
1-9
,共9页
马长江%王朝晖%杨雪%梁建新
馬長江%王朝暉%楊雪%樑建新
마장강%왕조휘%양설%량건신
微表层%浮游植物群落%DNA指纹%PCR-DGGE%大亚湾
微錶層%浮遊植物群落%DNA指紋%PCR-DGGE%大亞灣
미표층%부유식물군락%DNA지문%PCR-DGGE%대아만
The sea surface microlayer%Phytoplankton community%DNA fingerprint%PCR-DGGE%Daya Bay
为了解海洋微表层浮游植物群落结构的日变化特征,于2013年12月3日清晨(6：00)、正午(12：00)、傍晚(18：00)采集了大亚湾海域3个站位微表层和次表层水样,利用PCR-变性梯度凝胶电泳技术(PCR-DGGE)和显微镜观察对浮游植物DNA指纹及群落结构进行了分析比较。共分析鉴定出浮游植物79种,微表层和次表层浮分别为61种、68种,清晨、正午和傍晚观察的种类数分别为49种、61种、51种；清晨、正午和傍晚的平均细胞密度分别为1.2×106、1.6×106、1.6×106个/L,微表层和次表层平均细胞密度分别为1.72×106和1.22×106个/L。硅藻占有绝对优势,硅藻的数量百分比均在98%以上,主要优势硅藻为角毛藻(Chaetocerosspp.)、中肋骨条藻(Skeletonema costatum)、丹麦细柱藻(Leptocylindrus danicus)等。微表层对总浮游植物、硅藻及优势硅藻具有明显富集作用,其中对中肋骨条藻和丹麦细柱藻的富集系数分别为4.20和5.47,富集率均为100%。浮游植物DNA指纹条带也较丰富,每个样品DNA指纹条带数为12~28条,其中傍晚指纹条带最为丰富。由于优势种类对非优势种的屏蔽作用, DNA指纹条带数低于浮游植物种类数,但在剔除数量上小于0.5%的非优势种后, DNA指纹条带数与浮游植物种类数相近,说明PCR-DGGE技术对浮游植物检测灵敏度为优势度0.5%左右。
為瞭解海洋微錶層浮遊植物群落結構的日變化特徵,于2013年12月3日清晨(6：00)、正午(12：00)、傍晚(18：00)採集瞭大亞灣海域3箇站位微錶層和次錶層水樣,利用PCR-變性梯度凝膠電泳技術(PCR-DGGE)和顯微鏡觀察對浮遊植物DNA指紋及群落結構進行瞭分析比較。共分析鑒定齣浮遊植物79種,微錶層和次錶層浮分彆為61種、68種,清晨、正午和傍晚觀察的種類數分彆為49種、61種、51種；清晨、正午和傍晚的平均細胞密度分彆為1.2×106、1.6×106、1.6×106箇/L,微錶層和次錶層平均細胞密度分彆為1.72×106和1.22×106箇/L。硅藻佔有絕對優勢,硅藻的數量百分比均在98%以上,主要優勢硅藻為角毛藻(Chaetocerosspp.)、中肋骨條藻(Skeletonema costatum)、丹麥細柱藻(Leptocylindrus danicus)等。微錶層對總浮遊植物、硅藻及優勢硅藻具有明顯富集作用,其中對中肋骨條藻和丹麥細柱藻的富集繫數分彆為4.20和5.47,富集率均為100%。浮遊植物DNA指紋條帶也較豐富,每箇樣品DNA指紋條帶數為12~28條,其中傍晚指紋條帶最為豐富。由于優勢種類對非優勢種的屏蔽作用, DNA指紋條帶數低于浮遊植物種類數,但在剔除數量上小于0.5%的非優勢種後, DNA指紋條帶數與浮遊植物種類數相近,說明PCR-DGGE技術對浮遊植物檢測靈敏度為優勢度0.5%左右。
위료해해양미표층부유식물군락결구적일변화특정,우2013년12월3일청신(6：00)、정오(12：00)、방만(18：00)채집료대아만해역3개참위미표층화차표층수양,이용PCR-변성제도응효전영기술(PCR-DGGE)화현미경관찰대부유식물DNA지문급군락결구진행료분석비교。공분석감정출부유식물79충,미표층화차표층부분별위61충、68충,청신、정오화방만관찰적충류수분별위49충、61충、51충；청신、정오화방만적평균세포밀도분별위1.2×106、1.6×106、1.6×106개/L,미표층화차표층평균세포밀도분별위1.72×106화1.22×106개/L。규조점유절대우세,규조적수량백분비균재98%이상,주요우세규조위각모조(Chaetocerosspp.)、중륵골조조(Skeletonema costatum)、단맥세주조(Leptocylindrus danicus)등。미표층대총부유식물、규조급우세규조구유명현부집작용,기중대중륵골조조화단맥세주조적부집계수분별위4.20화5.47,부집솔균위100%。부유식물DNA지문조대야교봉부,매개양품DNA지문조대수위12~28조,기중방만지문조대최위봉부。유우우세충류대비우세충적병폐작용, DNA지문조대수저우부유식물충류수,단재척제수량상소우0.5%적비우세충후, DNA지문조대수여부유식물충류수상근,설명PCR-DGGE기술대부유식물검측령민도위우세도0.5%좌우。
Water samples from the sea surface microlayer (SML) and subsurface water (SSW) were collected from Daya Bay, the South China Sea at early morning (6: 00), mid noon (12: 00) and late afternoon (18: 00) on December 3rd 2013. The purpose is to understand daily changes of phytoplankton community in the SML. DNA fingerprints of phytoplankton were analyzed using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) technique, and phytoplankton was observed under the light microscope. A total of 79 taxa were identified during the survey in one day, including 61 species from the SML and 68 species from the SSW, and 49 species in the early morning, 61 species in the mid noon and 51 species in the late afternoon, respectively, with the average cells densities of 1.2×106cells/L, 1.6×106cells/L and 1.6×106cells/L, respectively, and 1.72×106 cells/Lin the SML and 1.22×106cells/L in the SSW. The diatoms were the preponderant group, the percentage compositions of diatoms were more than 98%, and the dominant diatom species includedChaetocerosspp., Skeletonema costatum andLeptocylindrus danicus et al. Diatoms and the overall phytoplankton were enriched in the SML, particular for the dominant diatom speciesS. costatum andL. danicus with enrichment factors of 4.20 and 5.47 and enrich-ment frequency of 100%, respectively. The DNA fingerprints were rich, and ranged from 12 to 28 for each sample. The fin-gerprint number was the highest in the late afternoon. The fingerprint numbers were lower than the species numbers observed under the microscope as the dominant species may screen the minority group during the PCR-DGGE process. However, they showed few differences after excluding those species less than 0.5% quantitatively. The results suggested that the DNA finger-prints using the PCR-DGGE technique might represent phytoplankton community with sensitivity about 0.5%.
