甘蔗斑茅杂交后代染色体遗传分析及品质抗性鉴定
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摘要
斑茅是甘蔗的近缘属植物之一,具有许多优异的性状,如抗旱、抗病、适应性广等多种优点,是甘蔗育种中具有较大潜在利用价值的重要资源。我国有丰富的斑茅资源,为了更好的利用斑茅资源,本文通过对斑茅和甘蔗杂交后代F_1、BC_1和BC_2进行了杂种真实性的ITS检验,并通过细胞遗传学对斑茅的BC_1和BC_2染色体遗传特性进行了初步分析,并对斑茅和甘蔗的杂种进行了耐旱性、抗黑穗病性和品质性状进行鉴定。主要结果以下:
(1)崖城01-69、崖城01-134、崖城01-116、崖城01-36、崖城03-06及崖城05-179在2条引物都出现了斑茅特异条带,这些供试材料都是甘蔗斑茅的真实杂种。崖城01-21在EF_1和ER_1中扩增出了特异条带,但在EF_2和ER_2中未扩增出条带,因此尚难断定崖城01-21是否为斑茅的真实杂种。结合染色体分析认为该材料也是斑茅真实杂种后代。
(2)核型分析研究表明:崖城01-69核型公式为2n=94=94m,染色体属1B型;崖城01-134核型公式为2n=114=114m,染色体属1B型;崖城01-116核型公式为2n=92=90m+2sm,染色体属1B型;崖城01-21核型公式为2n=104=100m+4sm(4SAT),染色体属2C型;崖城01-36为2n=132,染色体属2B型,核型公式为2n=132=130m+2sm;崖城03-06核型公式为2n=108=106m+2sm,染色体属2B型;崖城05-179为核型公式为2n=110=110m,染色体属1B型。内江57-416核型公式为2n=110=98m+12sm,染色体属2B型。甘蔗斑茅杂交后代的BC_1和BC_2中,崖城01-36染色体以2n+n的配对方式进行遗传,其余的以n+n的配对方式进行遗传,并且杂种染色体配对出现了不平衡的现象。
(3)各材料光合作用参数对水分胁迫的响应分为三个阶段。只是不同基因型的反应时间不同。在干旱胁迫下,植物气孔关闭,使CO_2摄取量减小,光合作用迅速下降。斑茅杂种后代崖城96-40在同等断水情况下,其光合能力、蒸腾速率、气孔导度都比其亲本及其它材料高,且Pn、Tr、Gs、Ci四个指标的变化趋势相同,表明Pn的下降还主要是以气孔因素为主。因此可认为崖城96-40的抗旱能力较强。严重水分胁迫下,供试材料中拔地拉及崖城01-03的MDA含量、质膜透性增加幅度较大,而海南92-105、海南92-77及斑茅F1崖城96-66的MDA含量、质膜透性增幅较小,崖城01-21与崖城96-40增幅低于CP84-1198或相当,说明斑茅及其后代具较强的耐旱性,但并不是所有斑茅后代都能遗传到斑茅的优良抗旱性状。抗旱性较强的品种能维持较高的净光合速率。
(4)崖城96-66、崖城01-36、崖城96-40和崖城01-21都高抗甘蔗黑穗病。斑茅能较好地把抗黑穗病性状遗传给后代,因此,它将成为甘蔗抗黑穗病性育种的一个重要抗源。斑茅及其F_1的蔗糖分含量很低,但经过回交改良后能显著提高,其遗传方式近似中亲遗传,这可能与染色体遗传方式相关。
Erianthus arundinaceus,with many outstanding traits such as drought tolerance,disease resistance and superiority adaptability,is a related genera plant of Saccharum L.It is also a promising germplasm for sugarcane breeding.There is abundant germplasm of E. Arundinaceus in China.In order to make better utilization the germplasm,we carried out the experiments of the calculation of chromosome number,karyotype analysis and ITS marker for hybrid,identification of drought tolerance and disease resistance and sucrose content analysis on the BC_1 and BC_2 of S.officinarum L.and E.Arundinaceus.
The results of ITS marker showed that the bands,with sizes of 250～500bp by two primer combinations of EF_1/ER_1 and EF_2/ER_2,specific to E.arundinaceus was observed in six genotypes of YC01-69,YC01 - 134,YC01-116,YC01-36,YC03-06 and YC05-179.It suggests that all of them were genuine progenies of E.arundinaceus and S.officinarum.Some differences were observed in YC01-21 when amplified by primer combinations of EF_1/ER_1 and EF_2/ER_2.For EF_1ER_1,the specific band was observed but not for EF_2/ER_2.YC 01-21 was at last identified as the progeny of E.arundinaceus by karyotypes analysis.
The results of chromosome number calculation and karyotype analysis show that the somatic chromosomes number of Neijiang57-416 is 2n= 110=98m+12sm and belongs to 2B type,Ya01-69 is 2n=94=94m and belongs to 1B type,Ya01-134 is 2n=114=114m and belongs to 1B type,Ya01-116 is 2n=92=90m+2sm and belongs to 1B type,Ya01-21 is 2n=104=100m+4sm(4SAT)and belongs to 2C type,Ya01-36 is 2n=132=130m+2sm and belongs to 1B type,Ya03-06 is 2n= 108= 106m+2sm and belongs to 2B type,Ya05-179 is 2n =110=110m and belongs to 1B type.The progenies BC_1 and BC_2 of E.arundinaceum and S. officinarum always follow the chromosome genetic law of n+n,only one of the progenies of E. arundinaceum and S.officinarum named YC01-36 follows 2n+n and the phenomena of 'not balance' chromosome pairing being observed.
The responses of photosynthetic parameters to drought stress are divided into three phases and difference in reaction time for difference genotypes.The reactions include stoma close up, less co_2 taking and photosynthesis ability decrease under drought stress.In the same condition, photosynthesis parameters of Pn,Tr,Gs and Ci for one of the progenies of E.arundinaceum YC96-40 are highest in all tested genotypes including its parents and the other progenies.And the change of these four parameters appeared the similar trends.It indicates the decrease of Pn is mainly due to the close up of stoma.It suggests YC96-40 with strong drought tolerance. Under the serious drought stress,Badila and YC01-03 increased greatly in the content of MDA and plasma membrane permeability(PMP)compared to the other genotypes.The smaller increment in the contents of MDA and PMP were observed in HN105,HN92-77 and YC96-66,one of F_1 material of E.arundinaceum.The increments of MDA and PMP in YC01-21 and YC96-40 were less than or equal to CP84-1198.It suggests E.arundinaceum and its progenies have a strong drought tolerance,but not all hybrid progenies will inherit this character.
YC96-66,YC01—36,YC96-40 and YC01-21 are highly resistant to sugarcane smut caused by Ustilago scitaminea Syd.This suggests that smut resistance for E.arundinaceum can transmitted to its progenies.Thus,E.arundinaceum will be an important resistance source in sugarcane breeding.Low sucrose content is observed in E.arundinaceum and its F_1 hybrids, but it can be significantly improved by backcross modification.The heredity of sucrose content character appears to be near mid-parent heredity.It may be related to the mode of chromosome heredity.
(1)崖城01-69、崖城01-134、崖城01-116、崖城01-36、崖城03-06及崖城05-179在2条引物都出现了斑茅特异条带,这些供试材料都是甘蔗斑茅的真实杂种。崖城01-21在EF_1和ER_1中扩增出了特异条带,但在EF_2和ER_2中未扩增出条带,因此尚难断定崖城01-21是否为斑茅的真实杂种。结合染色体分析认为该材料也是斑茅真实杂种后代。
(2)核型分析研究表明:崖城01-69核型公式为2n=94=94m,染色体属1B型;崖城01-134核型公式为2n=114=114m,染色体属1B型;崖城01-116核型公式为2n=92=90m+2sm,染色体属1B型;崖城01-21核型公式为2n=104=100m+4sm(4SAT),染色体属2C型;崖城01-36为2n=132,染色体属2B型,核型公式为2n=132=130m+2sm;崖城03-06核型公式为2n=108=106m+2sm,染色体属2B型;崖城05-179为核型公式为2n=110=110m,染色体属1B型。内江57-416核型公式为2n=110=98m+12sm,染色体属2B型。甘蔗斑茅杂交后代的BC_1和BC_2中,崖城01-36染色体以2n+n的配对方式进行遗传,其余的以n+n的配对方式进行遗传,并且杂种染色体配对出现了不平衡的现象。
(3)各材料光合作用参数对水分胁迫的响应分为三个阶段。只是不同基因型的反应时间不同。在干旱胁迫下,植物气孔关闭,使CO_2摄取量减小,光合作用迅速下降。斑茅杂种后代崖城96-40在同等断水情况下,其光合能力、蒸腾速率、气孔导度都比其亲本及其它材料高,且Pn、Tr、Gs、Ci四个指标的变化趋势相同,表明Pn的下降还主要是以气孔因素为主。因此可认为崖城96-40的抗旱能力较强。严重水分胁迫下,供试材料中拔地拉及崖城01-03的MDA含量、质膜透性增加幅度较大,而海南92-105、海南92-77及斑茅F1崖城96-66的MDA含量、质膜透性增幅较小,崖城01-21与崖城96-40增幅低于CP84-1198或相当,说明斑茅及其后代具较强的耐旱性,但并不是所有斑茅后代都能遗传到斑茅的优良抗旱性状。抗旱性较强的品种能维持较高的净光合速率。
(4)崖城96-66、崖城01-36、崖城96-40和崖城01-21都高抗甘蔗黑穗病。斑茅能较好地把抗黑穗病性状遗传给后代,因此,它将成为甘蔗抗黑穗病性育种的一个重要抗源。斑茅及其F_1的蔗糖分含量很低,但经过回交改良后能显著提高,其遗传方式近似中亲遗传,这可能与染色体遗传方式相关。
Erianthus arundinaceus,with many outstanding traits such as drought tolerance,disease resistance and superiority adaptability,is a related genera plant of Saccharum L.It is also a promising germplasm for sugarcane breeding.There is abundant germplasm of E. Arundinaceus in China.In order to make better utilization the germplasm,we carried out the experiments of the calculation of chromosome number,karyotype analysis and ITS marker for hybrid,identification of drought tolerance and disease resistance and sucrose content analysis on the BC_1 and BC_2 of S.officinarum L.and E.Arundinaceus.
The results of ITS marker showed that the bands,with sizes of 250～500bp by two primer combinations of EF_1/ER_1 and EF_2/ER_2,specific to E.arundinaceus was observed in six genotypes of YC01-69,YC01 - 134,YC01-116,YC01-36,YC03-06 and YC05-179.It suggests that all of them were genuine progenies of E.arundinaceus and S.officinarum.Some differences were observed in YC01-21 when amplified by primer combinations of EF_1/ER_1 and EF_2/ER_2.For EF_1ER_1,the specific band was observed but not for EF_2/ER_2.YC 01-21 was at last identified as the progeny of E.arundinaceus by karyotypes analysis.
The results of chromosome number calculation and karyotype analysis show that the somatic chromosomes number of Neijiang57-416 is 2n= 110=98m+12sm and belongs to 2B type,Ya01-69 is 2n=94=94m and belongs to 1B type,Ya01-134 is 2n=114=114m and belongs to 1B type,Ya01-116 is 2n=92=90m+2sm and belongs to 1B type,Ya01-21 is 2n=104=100m+4sm(4SAT)and belongs to 2C type,Ya01-36 is 2n=132=130m+2sm and belongs to 1B type,Ya03-06 is 2n= 108= 106m+2sm and belongs to 2B type,Ya05-179 is 2n =110=110m and belongs to 1B type.The progenies BC_1 and BC_2 of E.arundinaceum and S. officinarum always follow the chromosome genetic law of n+n,only one of the progenies of E. arundinaceum and S.officinarum named YC01-36 follows 2n+n and the phenomena of 'not balance' chromosome pairing being observed.
The responses of photosynthetic parameters to drought stress are divided into three phases and difference in reaction time for difference genotypes.The reactions include stoma close up, less co_2 taking and photosynthesis ability decrease under drought stress.In the same condition, photosynthesis parameters of Pn,Tr,Gs and Ci for one of the progenies of E.arundinaceum YC96-40 are highest in all tested genotypes including its parents and the other progenies.And the change of these four parameters appeared the similar trends.It indicates the decrease of Pn is mainly due to the close up of stoma.It suggests YC96-40 with strong drought tolerance. Under the serious drought stress,Badila and YC01-03 increased greatly in the content of MDA and plasma membrane permeability(PMP)compared to the other genotypes.The smaller increment in the contents of MDA and PMP were observed in HN105,HN92-77 and YC96-66,one of F_1 material of E.arundinaceum.The increments of MDA and PMP in YC01-21 and YC96-40 were less than or equal to CP84-1198.It suggests E.arundinaceum and its progenies have a strong drought tolerance,but not all hybrid progenies will inherit this character.
YC96-66,YC01—36,YC96-40 and YC01-21 are highly resistant to sugarcane smut caused by Ustilago scitaminea Syd.This suggests that smut resistance for E.arundinaceum can transmitted to its progenies.Thus,E.arundinaceum will be an important resistance source in sugarcane breeding.Low sucrose content is observed in E.arundinaceum and its F_1 hybrids, but it can be significantly improved by backcross modification.The heredity of sucrose content character appears to be near mid-parent heredity.It may be related to the mode of chromosome heredity.
引文
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