分子植物育种
分子植物育種
분자식물육충
MOLECULAR PLANT BREEDING
2013年
4期
509-516
,共8页
苏顺宗%刘丹%吴玲%聂治%张啸%易双%高世斌
囌順宗%劉丹%吳玲%聶治%張嘯%易雙%高世斌
소순종%류단%오령%섭치%장소%역쌍%고세빈
玉米%ZmPAP18%低磷胁迫%表达分析%序列变异
玉米%ZmPAP18%低燐脅迫%錶達分析%序列變異
옥미%ZmPAP18%저린협박%표체분석%서렬변이
Zea mays%ZmPAP18%Low phosphorus stress%Expression analysis%Sequence variation
紫色酸性磷酸酶(PAP)是金属磷酸酯酶家族的重要成员,在植物对低磷环境适应的过程中发挥重要作用。依据前期构建的低磷胁迫抑制消减文库,运用RT-PCR方法从耐低磷玉米自交系178中克隆了一个酸性磷酸酶基因的全长序列。该基因编码区全长1409 bp,其中ORF为1371 bp,编码457个氨基酸残基。经保守域分析发现,基因的推测氨基酸序列中含有紫色酸性磷酸酶所特有的5个保守基序和7个金属离子结合位点,属于紫色酸性磷酸酶类,被命名为ZmPAP18。通过实时荧光定量分析表明,ZmPAP18在根和叶中均受到低磷胁迫的诱导增强表达,且在诱导24 h后表达量达到峰值;但在玉米根部的表达量高于叶片,表明ZmPAP18在玉米根部可能参与了根外磷素的活化与吸收以及无机磷由根部向其他组织的转运。此外,经序列分析发现,ZmPAP18的3'非翻译区存在一个Indel (TGTTG)位点,并在常用的14个玉米自交系中表现出丰富的多态性,为进一步开发该基因的分子标记及耐低磷种质分子改良提供了参考信息。
紫色痠性燐痠酶(PAP)是金屬燐痠酯酶傢族的重要成員,在植物對低燐環境適應的過程中髮揮重要作用。依據前期構建的低燐脅迫抑製消減文庫,運用RT-PCR方法從耐低燐玉米自交繫178中剋隆瞭一箇痠性燐痠酶基因的全長序列。該基因編碼區全長1409 bp,其中ORF為1371 bp,編碼457箇氨基痠殘基。經保守域分析髮現,基因的推測氨基痠序列中含有紫色痠性燐痠酶所特有的5箇保守基序和7箇金屬離子結閤位點,屬于紫色痠性燐痠酶類,被命名為ZmPAP18。通過實時熒光定量分析錶明,ZmPAP18在根和葉中均受到低燐脅迫的誘導增彊錶達,且在誘導24 h後錶達量達到峰值;但在玉米根部的錶達量高于葉片,錶明ZmPAP18在玉米根部可能參與瞭根外燐素的活化與吸收以及無機燐由根部嚮其他組織的轉運。此外,經序列分析髮現,ZmPAP18的3'非翻譯區存在一箇Indel (TGTTG)位點,併在常用的14箇玉米自交繫中錶現齣豐富的多態性,為進一步開髮該基因的分子標記及耐低燐種質分子改良提供瞭參攷信息。
자색산성린산매(PAP)시금속린산지매가족적중요성원,재식물대저린배경괄응적과정중발휘중요작용。의거전기구건적저린협박억제소감문고,운용RT-PCR방법종내저린옥미자교계178중극륭료일개산성린산매기인적전장서렬。해기인편마구전장1409 bp,기중ORF위1371 bp,편마457개안기산잔기。경보수역분석발현,기인적추측안기산서렬중함유자색산성린산매소특유적5개보수기서화7개금속리자결합위점,속우자색산성린산매류,피명명위ZmPAP18。통과실시형광정량분석표명,ZmPAP18재근화협중균수도저린협박적유도증강표체,차재유도24 h후표체량체도봉치;단재옥미근부적표체량고우협편,표명ZmPAP18재옥미근부가능삼여료근외린소적활화여흡수이급무궤린유근부향기타조직적전운。차외,경서렬분석발현,ZmPAP18적3'비번역구존재일개Indel (TGTTG)위점,병재상용적14개옥미자교계중표현출봉부적다태성,위진일보개발해기인적분자표기급내저린충질분자개량제공료삼고신식。
Purple acid phosphatase (PAP) is a gene family of metallo-phosphoesterases involved in a variety of physiological functions, especially phosphate (Pi) deficiency adaptations in plants. Based on the cDNA library constructed by SSH for maize seedling root under phosphorus deficiency. A 1 409 bp length gene was cloned from low-phosphate tolerant maize inbred line 178, which encoded 457 amino acid residues. The conserved domain analysis revealed that this cloned gene had five conserved motifs and seven invariant metal-ligating residues typical of known PAPs and therefore named as ZmPAP18. Expression profiles of ZmPAP18 were assayed by fluorescent quantitative PCR and indicated that ZmPAP18 was up-regulated both in seedling root and leaf under phosphate starvation. The expression levels increased in the early stage of Pi starvation, and then continuously increased to the maximal level at 24 h time point. In comparison with seedling leaf, ZmPAP18 was more sensitive and responsive to phosphate starvation in seedling root. These results suggested that ZmPAP18 in maize may be involved in external phosphorus assimilation and transferring inorganic phosphate from root to other tissues. Moreover, an Indel (TGTTG) site was detected in 3' UTR of ZmPAP18 and showed highly polymorphism among 14 maize inbred lines, which can be developed as molecular marker to assist improving P utilization in molecular maize breeding.