植物营养与肥料学报
植物營養與肥料學報
식물영양여비료학보
PLANT NUTRITION AND FERTILIZER SCIENCE
2014年
4期
877-884
,共8页
郭丽%郭程瑾%路文静%李小娟%肖凯
郭麗%郭程瑾%路文靜%李小娟%肖凱
곽려%곽정근%로문정%리소연%초개
小麦( Triticum aestivum L. )%磷转运蛋白基因%低磷胁迫%磷素吸收%干物质生产
小麥( Triticum aestivum L. )%燐轉運蛋白基因%低燐脅迫%燐素吸收%榦物質生產
소맥( Triticum aestivum L. )%린전운단백기인%저린협박%린소흡수%간물질생산
wheat ( Triticum aestivum L. )%phosphate transporter gene%Pi deprivation%phosphate acquisition%dry mass production
【目的】植株对介质中磷素的吸收及磷素在体内器官组织间的转运,是通过位于细胞质膜上的磷转运蛋白( PT)介导完成的。高亲和PT在介导植物对低磷逆境下的磷素吸收中发挥重要作用。本研究以小麦中国春遗传背景的整套B染色体双端体为材料,对小麦高亲和PT基因TaPht1；4的染色体定位特征及其与低磷下小麦品种磷效率的联系进行系统研究,旨在为今后小麦品种磷效率分子鉴定和磷高效遗传改良提供依据。【方法】采用水培法培养中国春( CS)及其遗传背景B染色体组双端体幼苗。三叶期时收获各供试材料根系,提取各材料基因组DNA,通过PCR特异扩增TaPht1；4,鉴定TaPht1；4在染色体上定位。通过对各供试材料三叶期幼苗进行24 h低磷胁迫获取丰缺磷处理根叶样本,采用半定量RT-PCR及实时定量PCR分析TaPht1；4在丰缺磷下的表达。采用上述幼苗培养、丰缺磷处理和基因表达分析技术,研究不同磷吸收效率小麦品种磷效率参数和TaPht1；4表达特征。【结果】1)与CS及其他双端体材料能特异扩增目标基因不同,在3BS中未扩增到目标基因TaPht1；4；采用半定量RT-PCR和qPCR对丰、缺磷下CS和各双端体根、叶中TaPht1；4的表达研究表明,丰磷下各供试材料根、叶中均检测不到TaPht1；4表达,缺磷下各供试材料叶片中也均未检测到 TaPht1；4表达,但在根中除3BS 未检测到TaPht1；4表达外,CS和其他双端体均具有较高的TaPht1；4表达水平。表明TaPht1；4定位在3B染色体长臂,呈低磷诱导和根系特异表达特征。2)丰磷下,3BS单株干重与CS没有差异；缺磷下,与CS相比,3BS单株干重显著降低。表明缺少TaPht1；4及所在3B染色体长臂后,植株干物质生产能力受到较大影响,这可能与因缺乏该染色体臂丧失TaPht1；4造成低磷下植株的磷素吸收能力降低密切相关。3)对丰、缺磷下不同磷吸收效率6个小麦品种TaPht1；4的表达水平以及单株干重、全磷含量、磷累积量和磷效率研究表明,缺磷下各小麦品种表现为随品种磷吸收效率提高,TaPht1；4表达水平也随之增高。表明TaPht1；4表达水平与低磷下小麦品种磷素吸收能力和干物质积累具有紧密联系。【结论】小麦高亲和PT基因TaPht1；4定位在3B长臂。低磷条件下,3BS的单株干重和磷累积量较CS显著降低。丰、缺磷下,不同磷吸收效率小麦品种TaPht1；4表达水平与植株干重和单株磷累积量密切相关。 TaPht1；4能显著增强小麦在低磷下磷素吸收能力,可作为小麦品种耐低磷能力的参考分子评价指标。
【目的】植株對介質中燐素的吸收及燐素在體內器官組織間的轉運,是通過位于細胞質膜上的燐轉運蛋白( PT)介導完成的。高親和PT在介導植物對低燐逆境下的燐素吸收中髮揮重要作用。本研究以小麥中國春遺傳揹景的整套B染色體雙耑體為材料,對小麥高親和PT基因TaPht1；4的染色體定位特徵及其與低燐下小麥品種燐效率的聯繫進行繫統研究,旨在為今後小麥品種燐效率分子鑒定和燐高效遺傳改良提供依據。【方法】採用水培法培養中國春( CS)及其遺傳揹景B染色體組雙耑體幼苗。三葉期時收穫各供試材料根繫,提取各材料基因組DNA,通過PCR特異擴增TaPht1；4,鑒定TaPht1；4在染色體上定位。通過對各供試材料三葉期幼苗進行24 h低燐脅迫穫取豐缺燐處理根葉樣本,採用半定量RT-PCR及實時定量PCR分析TaPht1；4在豐缺燐下的錶達。採用上述幼苗培養、豐缺燐處理和基因錶達分析技術,研究不同燐吸收效率小麥品種燐效率參數和TaPht1；4錶達特徵。【結果】1)與CS及其他雙耑體材料能特異擴增目標基因不同,在3BS中未擴增到目標基因TaPht1；4；採用半定量RT-PCR和qPCR對豐、缺燐下CS和各雙耑體根、葉中TaPht1；4的錶達研究錶明,豐燐下各供試材料根、葉中均檢測不到TaPht1；4錶達,缺燐下各供試材料葉片中也均未檢測到 TaPht1；4錶達,但在根中除3BS 未檢測到TaPht1；4錶達外,CS和其他雙耑體均具有較高的TaPht1；4錶達水平。錶明TaPht1；4定位在3B染色體長臂,呈低燐誘導和根繫特異錶達特徵。2)豐燐下,3BS單株榦重與CS沒有差異；缺燐下,與CS相比,3BS單株榦重顯著降低。錶明缺少TaPht1；4及所在3B染色體長臂後,植株榦物質生產能力受到較大影響,這可能與因缺乏該染色體臂喪失TaPht1；4造成低燐下植株的燐素吸收能力降低密切相關。3)對豐、缺燐下不同燐吸收效率6箇小麥品種TaPht1；4的錶達水平以及單株榦重、全燐含量、燐纍積量和燐效率研究錶明,缺燐下各小麥品種錶現為隨品種燐吸收效率提高,TaPht1；4錶達水平也隨之增高。錶明TaPht1；4錶達水平與低燐下小麥品種燐素吸收能力和榦物質積纍具有緊密聯繫。【結論】小麥高親和PT基因TaPht1；4定位在3B長臂。低燐條件下,3BS的單株榦重和燐纍積量較CS顯著降低。豐、缺燐下,不同燐吸收效率小麥品種TaPht1；4錶達水平與植株榦重和單株燐纍積量密切相關。 TaPht1；4能顯著增彊小麥在低燐下燐素吸收能力,可作為小麥品種耐低燐能力的參攷分子評價指標。
【목적】식주대개질중린소적흡수급린소재체내기관조직간적전운,시통과위우세포질막상적린전운단백( PT)개도완성적。고친화PT재개도식물대저린역경하적린소흡수중발휘중요작용。본연구이소맥중국춘유전배경적정투B염색체쌍단체위재료,대소맥고친화PT기인TaPht1；4적염색체정위특정급기여저린하소맥품충린효솔적련계진행계통연구,지재위금후소맥품충린효솔분자감정화린고효유전개량제공의거。【방법】채용수배법배양중국춘( CS)급기유전배경B염색체조쌍단체유묘。삼협기시수획각공시재료근계,제취각재료기인조DNA,통과PCR특이확증TaPht1；4,감정TaPht1；4재염색체상정위。통과대각공시재료삼협기유묘진행24 h저린협박획취봉결린처리근협양본,채용반정량RT-PCR급실시정량PCR분석TaPht1；4재봉결린하적표체。채용상술유묘배양、봉결린처리화기인표체분석기술,연구불동린흡수효솔소맥품충린효솔삼수화TaPht1；4표체특정。【결과】1)여CS급기타쌍단체재료능특이확증목표기인불동,재3BS중미확증도목표기인TaPht1；4；채용반정량RT-PCR화qPCR대봉、결린하CS화각쌍단체근、협중TaPht1；4적표체연구표명,봉린하각공시재료근、협중균검측불도TaPht1；4표체,결린하각공시재료협편중야균미검측도 TaPht1；4표체,단재근중제3BS 미검측도TaPht1；4표체외,CS화기타쌍단체균구유교고적TaPht1；4표체수평。표명TaPht1；4정위재3B염색체장비,정저린유도화근계특이표체특정。2)봉린하,3BS단주간중여CS몰유차이；결린하,여CS상비,3BS단주간중현저강저。표명결소TaPht1；4급소재3B염색체장비후,식주간물질생산능력수도교대영향,저가능여인결핍해염색체비상실TaPht1；4조성저린하식주적린소흡수능력강저밀절상관。3)대봉、결린하불동린흡수효솔6개소맥품충TaPht1；4적표체수평이급단주간중、전린함량、린루적량화린효솔연구표명,결린하각소맥품충표현위수품충린흡수효솔제고,TaPht1；4표체수평야수지증고。표명TaPht1；4표체수평여저린하소맥품충린소흡수능력화간물질적루구유긴밀련계。【결론】소맥고친화PT기인TaPht1；4정위재3B장비。저린조건하,3BS적단주간중화린루적량교CS현저강저。봉、결린하,불동린흡수효솔소맥품충TaPht1；4표체수평여식주간중화단주린루적량밀절상관。 TaPht1；4능현저증강소맥재저린하린소흡수능력,가작위소맥품충내저린능력적삼고분자평개지표。
[Objectives]The acquisition of inorganic phosphate ( Pi) and the Pi translocation across the organs and tissues in plants is mediated by phosphate transporters ( PTs) located at the cytoplasmic membranes. The PTs with high-affinity property play critical roles in mediating the Pi absorption by plants under the Pi-limited condition. Currently, the molecular characterization and biological functions of PTs in wheat were few reported. In this study, using Chinese spring ( CS) and its ditelosimic lines of B chromosome as materials, the localization on chromosome as well as expression patterns of TaPht1; 4, a high-affinity PT gene in wheat, was systematically studied under sufficient-and deficient-Pi conditions. In addition, the relationship between the expression levels of TaPht1;4 and the plant phosphorus use efficiencies across various wheat cultivars under supply of lower phosphorous was determined to provide molecular basis for evaluation of phosphorus use efficiency across wheat cultivars and guidance on genetic improvement for high phosphorus usage in wheat.[Methods]The hydroponic culture method was used to cultivate seedlings of Chinese spring ( CS) and its ditelosimic lines of B chromosome. The roots and leaves of all tested materials were separately harvested at three-leaf growth stage and subjected to extraction of genome DNA. PCR analysis was performed using the gene specific primers of TaPht1;4 to identify the chromosome location of TaPht1;4. The roots and leaves of all tested materials under conditions of sufficient-and deficient-Pi by treated with 24 h low-Pi stress were sampled and subjected to expression analysis of TaPht1; 4 gene by semiquantitative RT-PCR and real-time PCR. Using the similar methods of seedling culture, treatments of sufficient-and deficient-Pi, and gene expression analysis approaches, the parameters of phosphorus use efficiency and the expression patterns of TaPht1;4 in wheat cultivars were investigated.[Results] 1) In contrast to CS and other its ditelosimic lines of B chromosome, TaPht1;4 was not amplified by PCR in 3BS. Moreover, the expression analysis of TaPht1;4 by semiquantitative RT-PCR and real-time PCR revealed that CS and all ditelosimic lines of B chromosome both in the roots and leaves under the condition of sufficient- Pi did not exhibit the expression of TaPht1;4, and similar result was shown in the leaves under the condition of deficient-Pi. However, the transcripts of TaPht1;4 were detected in the roots of CS as well as in the ditelosimic lines, and was not detected in that of 3BS. Therefore, TaPht1;4 is located at the long arm of 3B and exhibits expression patterns of induction by low-Pi stress, and has root-specific expression. 2)Under the condition of sufficient-Pi, no variations on plant dry mass was observed between the 3BS and CS. By contrast, the plant dry mass of 3BS was significantly lower than that of CS under the condition of deficient-Pi. These results indicate that the plant dry matter production is largely affected by TaPht1;4 and the long arm of 3 B that situates in the TaPht1; 4. Deletion of TaPht1; 4 by missing this chromosome arm can result in significant reduction in plant dry mass under deprivation of Pi. 3 ) Investigations on the expression levels of TaPht1;4,the plant dry mass, total phosphorus contents, accumulative phosphorus amount per plant, and phosphorus use efficiencies in six wheat cultivars with contrasting Pi acquisition capacity indicated that the expression levels of TaPht1;4 was positively correlated with the Pi acquisition capacities and phosphorus use efficiencies under the condition of deficient-Pi.[Conclusions]The wheat high affinity PT gene, TaPht1;4 is located at the long arm of 3B. Under the condition of deficient-Pi, the plant dry mass and accumulative amounts of phosphorus per plant of 3BS were significantly lower than those of CS. The expression levels of TaPht1;4 across the wheat cultivars with contrasting Pi acquisition capacity were closely associated with their plant dry masses and accumulative amounts of phosphorus per plant under conditions of sufficient- and deficient-Pi. Taken together, TaPht1;4 plays critical roles in regulating plant Pi acquisition in wheat under low-Pi stress, and can act as one of valuable molecular criteria in evaluating the capacity to tolerate low-Pi stress in wheat.
