近日,,國際雜志Molecular Ecology Resources刊登了中科院昆明動(dòng)物所車靜副研究員等的最新研究成果“Universal COI primers for DNA barcoding amphibians,。”,,文章中,研究者在兩棲類DNA條形碼通用引物研究方面取得了新的進(jìn)展,。
DNA條形碼技術(shù)(DNA Barcoding)近年來已成為國際上生物多樣性研究的熱點(diǎn),即利用標(biāo)準(zhǔn)基因片段對物種進(jìn)行快速、準(zhǔn)確的鑒定和識(shí)別,。該技術(shù)正廣泛應(yīng)用于生物多樣性調(diào)查、分子進(jìn)化,、生態(tài),、環(huán)境、食品安全等諸多領(lǐng)域,。
在脊椎動(dòng)物中,,目前魚類,鳥類,,以及極地動(dòng)物在DNA條形碼研究中進(jìn)展較快,,已有大量相關(guān)數(shù)據(jù)庫在線。作為重要類群之一的兩棲爬行動(dòng)物,,進(jìn)展一直較為緩慢,。在對兩棲類的研究中,歐洲學(xué)者探討了DNA條形碼的鑒別能力,,結(jié)果表明COI基因在該類群中鑒別物種方面是適合的,,但是由于COI基因進(jìn)化速率較快,序列保守性相對低,,引物通用性差,,極大阻礙了在全球兩棲類群中開展相關(guān)工作,。
中科院昆明動(dòng)物所車靜副研究員以及學(xué)生陳宏滿和楊軍校等人在張亞平院士的指導(dǎo)下,重新設(shè)計(jì)和優(yōu)化了兩對COI通用兼并引物,,在中國兩棲類的11個(gè)科,、36個(gè)屬、82個(gè)種111個(gè)個(gè)體中進(jìn)行了驗(yàn)證,,證實(shí)新設(shè)計(jì)的兩對引物能夠較廣泛的用于兩棲類DNA條形碼研究,。該結(jié)果目前已在線發(fā)表Molecular Ecology Resources。該引物目前在爬行類群測試中也得到很好的驗(yàn)證,。
兩棲爬行類是動(dòng)物界中獨(dú)特的一支,,尤其兩棲類代表由水到陸的轉(zhuǎn)型過渡物種,對環(huán)境變化極為敏感,,一直是研究環(huán)境和氣候變化的指示物種,。針對兩棲爬行類DNA條形碼工作不足的現(xiàn)狀,昆明動(dòng)物所于第四屆國際生命條形碼大會(huì)上提出兩棲爬行動(dòng)物DNA條形碼“Cold Code”計(jì)劃,,致力于在全球范圍內(nèi)領(lǐng)導(dǎo)兩棲爬行類標(biāo)準(zhǔn)標(biāo)記基因篩選,、技術(shù)體系構(gòu)建、標(biāo)準(zhǔn)文庫建設(shè)等方面發(fā)揮重要作用,。
本研究得到了科技部基礎(chǔ)性工作專項(xiàng),、基金委重大項(xiàng)目、中國科學(xué)院知識(shí)創(chuàng)新工程項(xiàng)目等的支持,。(生物谷Bioon.com)
doi:10.1111/j.1755-0998.2011.03090.x
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Universal COI primers for DNA barcoding amphibians
JING CHE1,*, HONG-MAN CHEN1,2,*, JUN-XIAO YANG1, JIE-QIONG JIN1, KE JIANG1, ZHI-YONG YUAN1, ROBERT W. MURPHY1,3, YA-PING ZHANG1,2
DNA barcoding is a proven tool for the rapid and unambiguous identification of species, which is essential for many activities including the vouchering tissue samples in the genome 10K initiative, genealogical reconstructions, forensics and biodiversity surveys, among many other applications. A large-scale effort is underway to barcode all amphibian species using the universally sequenced DNA region, a partial fragment of mitochondrial cytochrome oxidase subunit I COI. This fragment is desirable because it appears to be superior to 16S for barcoding, at least for some groups of salamanders. The barcoding of amphibians is essential in part because many species are now endangered. Unfortunately, existing primers for COI often fail to achieve this goal. Herein, we report two new pairs of primers (➀, ➁) that in combination serve to universally amplify and sequence all three orders of Chinese amphibians as represented by 36 genera. This taxonomic diversity, which includes caecilians, salamanders and frogs, suggests that the new primer pairs will universally amplify COI for the vast majority species of amphibians.