Aoki N, Scofield G N, Wang X-D, Patrick J W, Offler C E, Furbank R T
CSIRO Plant Industry, GPO Box 1600, ACT 2601, Canberra, Australia.
Planta. 2004 May;219(1):176-84. doi: 10.1007/s00425-004-1232-7. Epub 2004 Mar 11.
Previously we reported the isolation of three sucrose transporter genes, TaSUT1A, 1B and 1D, all expressed at high levels in the developing grains of hexaploid wheat ( Triticum aestivum L.), but also in a variety of other tissues. In order to further characterise the expression of the TaSUT1 genes in wheat plants, we have analysed TaSUT1 expression in their vegetative tissues using semi-quantitative reverse transcription-polymerase chain reaction, in situ hybridisation and immunolocalisation. The three TaSUT1 genes, which encode 98% identical SUT proteins, all appeared to be expressed at the same level in leaf blades, leaf sheaths and internodes, as well as developing grains, of hexaploid wheat. In mature leaf blades, TaSUT1 protein localised to the plasma membrane of phloem sieve elements in all classes of veins. In contrast, TaSUT1 mRNA was found to be localised to phloem companion cells. A similar localisation pattern for TaSUT1 protein was observed in veins of leaf sheaths and internodes. These results suggest that the wheat SUT1 has a transport function in enucleate sieve elements, in both veins responsible for loading photoassimilates, and in veins for axial transport. Furthermore, transport of the fluorescent dye carboxyfluorescein was used to investigate symplasmic connectivity between sieve element-companion cell complexes and non-phloem cells. Observations in source leaves indicated that sieve element-companion cell complexes of minor veins were symplasmically restricted, suggesting a role of TaSUT1 in apoplasmic phloem loading. In contrast, the dye was able to move symplasmically out of the phloem in internodes. In these circumstances TaSUT1 may also have a role in retrieving sucrose leaked to the phloem apoplasm.
此前我们报道了三个蔗糖转运蛋白基因TaSUT1A、1B和1D的分离,它们在六倍体小麦(Triticum aestivum L.)发育中的籽粒中高水平表达,在多种其他组织中也有表达。为了进一步表征TaSUT1基因在小麦植株中的表达情况,我们利用半定量逆转录-聚合酶链反应、原位杂交和免疫定位分析了TaSUT1在其营养组织中的表达。这三个TaSUT1基因编码的蔗糖转运蛋白有98%的同一性,在六倍体小麦的叶片、叶鞘、节间以及发育中的籽粒中,它们的表达水平似乎相同。在成熟叶片中,TaSUT1蛋白定位于所有类型叶脉中韧皮部筛管分子的质膜。相反,TaSUT1 mRNA定位于韧皮部伴胞。在叶鞘和节间的叶脉中也观察到了TaSUT1蛋白类似的定位模式。这些结果表明,小麦SUT1在无核筛管分子中具有转运功能,既在负责装载光合产物的叶脉中,也在负责轴向运输的叶脉中。此外,利用荧光染料羧基荧光素的运输来研究筛管分子-伴胞复合体与非韧皮部细胞之间的共质体连接性。在源叶中的观察结果表明,小叶脉的筛管分子-伴胞复合体在共质体上是受限的,这表明TaSUT1在质外体韧皮部装载中起作用。相反,染料能够在节间从韧皮部共质体中移出。在这种情况下,TaSUT1可能在回收泄漏到韧皮部质外体中的蔗糖方面也发挥作用。