Wiberley-Bradford Amy E, Busse James S, Jiang Jiming, Bethke Paul C
United States Department of Agriculture - Agricultural Research Service, Vegetable Crops Research Unit, 1575 Linden Dr,, Madison, WI 53706, USA.
BMC Res Notes. 2014 Nov 16;7:801. doi: 10.1186/1756-0500-7-801.
Storing potato tubers at low temperatures minimizes sprouting and disease but can cause an accumulation of reducing sugars in a process called cold-induced sweetening. Tubers with increased amounts of reducing sugars produce dark-colored, bitter-tasting fried products with elevated amounts of acrylamide, a possible carcinogen. Vacuolar invertase (VInv), which converts sucrose produced by starch breakdown to glucose and fructose, is the key determinant of reducing sugar accumulation during cold-induced sweetening. In this study, wild-type tubers and tubers in which VInv expression was reduced by RNA interference were used to investigate time- and temperature-dependent changes in sugar contents, chip color, and expression of VInv and other genes involved in starch metabolism in tubers during long-term cold storage.
VInv activities and tuber reducing sugar contents were much lower, and tuber sucrose contents were much higher, in transgenic than in wild-type tubers stored at 3-9°C for up to eight months. Large differences in VInv mRNA accumulation were not observed at later times in storage, especially at temperatures below 9°C, so differences in invertase activity were likely established early in the storage period and maintained by stability of the invertase protein. Sugar contents, chip color, and expression of several of the studied genes, including AGPase and GBSS, were affected by storage temperature in both wild-type and transgenic tubers. Though transcript accumulation for other sugar-metabolism genes was affected by storage temperature and duration, it was essentially unaffected by invertase silencing and altered sugar contents. Differences in stem- and bud-end sugar contents in wild-type and transgenic tubers suggested different compartmentalization of sucrose at the two ends of stored tubers.
VInv silencing significantly reduced cold-induced sweetening in stored potato tubers, likely by means of differential VInv expression early in storage. Transgenic tubers retained sensitivity to storage temperature, and accumulated greater amounts of sucrose, glucose and fructose at 3°C than at 7-9°C. At each storage temperature, suppression of VInv expression and large differences in tuber sugar contents had no effect on expression of AGPase and GBSS, genes involved in starch metabolism, suggesting that transcription of these genes is not regulated by tuber sugar content.
将马铃薯块茎低温储存可最大程度减少发芽和疾病,但在一个称为冷诱导糖化的过程中会导致还原糖积累。还原糖含量增加的块茎会产生颜色深、味道苦的油炸产品,且丙烯酰胺含量升高,丙烯酰胺是一种潜在致癌物。液泡转化酶(VInv)可将淀粉分解产生的蔗糖转化为葡萄糖和果糖,是冷诱导糖化过程中还原糖积累的关键决定因素。在本研究中,使用野生型块茎和通过RNA干扰降低VInv表达的块茎,来研究长期冷藏期间块茎中糖含量、薯片颜色以及VInv和其他参与淀粉代谢的基因表达随时间和温度的变化。
在3-9°C储存长达八个月的转基因块茎中,VInv活性和块茎还原糖含量远低于野生型块茎,而块茎蔗糖含量则远高于野生型块茎。在储存后期未观察到VInv mRNA积累有很大差异,尤其是在9°C以下的温度,因此转化酶活性的差异可能在储存期早期就已形成,并通过转化酶蛋白的稳定性得以维持。野生型和转基因块茎中的糖含量、薯片颜色以及包括AGPase和GBSS在内的几个研究基因的表达均受储存温度影响。尽管其他糖代谢基因的转录积累受储存温度和持续时间影响,但基本上不受转化酶沉默和糖含量变化的影响。野生型和转基因块茎茎端和芽端糖含量的差异表明,储存块茎两端的蔗糖存在不同的区室化。
VInv沉默显著降低了储存马铃薯块茎的冷诱导糖化,可能是通过储存早期VInv的差异表达实现的。转基因块茎对储存温度仍保持敏感性,在3°C时比在7-9°C时积累更多的蔗糖、葡萄糖和果糖。在每个储存温度下,VInv表达的抑制和块茎糖含量的巨大差异对参与淀粉代谢的基因AGPase和GBSS的表达没有影响,这表明这些基因的转录不受块茎糖含量的调节。