A 94-bp Deletion of Anthocyanidin Synthase Gene in Acyanic Flower Lines of Lisianthus [Eustoma grandiflorum (Raf.) Shinn.]

Abstract

We studied the genomic sequence of the anthocyanidin synthase (ANS) gene in acyanic flower lines of lisianthus and detected a 94-bp deletion of the sequence in comparison with that of cyanic flower lines. Cross-pollination was made between acyanic and cyanic flower lines to yield F 1 progeny, followed by its self-pollination. The segregated individuals between cyanic and acyanic flowers among the F 2 population were of 24 and 8, respectively (3 : 1), which suggested that one gene of the F 1 hybrid was unequivocally involved in anthocyanin production. The homozygous ANS genes that have a set of 94-bp deletion co-segregated a recessive phenotype of white color in the F 2 population. Genomic PCR of the ANS genes of the 14 independent lisianthus lines of acyanic flowers, such as white, yellow, pale yellow, and pale green flowers, revealed that all possessed a homozygous 94-bp deletion. The data support that the acyanic flower colors are associated with a 94-bp deletion in the ANS gene as a result of the loss of anthocyanin accumulation.

Recent advances on the development and regulation of flower color in ornamental plants

“…Studies have shown that ANS is encoded by a small gene family in many plants, and these genes have been cloned from ornamental plants, including Forsythia supensa ( Rosati et al, 1999 ), gerbera ( Wellmann et al, 2006 ) and herbaceous peony ( Zhao et al, 2012b ). Rosati et al (1999) studied the ANS gene expression pattern in Forsythia supensa and found that null expression of the ANS gene resulted in little accumulation of anthocyanins in petals; similarly, the absence of the ANS gene sequence was the underlying reason for the color change of lisianthus flowers ( Shimizu et al, 2011 ), suggesting its importance in the regulation of plant colors.…”

Evolution and developmental genetics of floral display—A review of progress

“…In nature, mutations causing loss of function in either the coding region or regulatory region of structural genes involved in the anthocyanin biosynthesis pathway were found to be the primarily cause for the evolutionary transition of flower colors in Petunia, Ipomoea, and Eustoma (Habu et al, 1998;Zufall & Rausher, 2004;Hoballah et al, 2007;Shimizu et al, 2011). Loss-of-function mutations often involve insertions or deletions in the coding region as a result of imprecise transposon exertion (Hoballah et al, 2007;Shimizu et al, 2011).…”

“…In nature, mutations causing loss of function in either the coding region or regulatory region of structural genes involved in the anthocyanin biosynthesis pathway were found to be the primarily cause for the evolutionary transition of flower colors in Petunia , Ipomoea , and Eustoma (Habu et al, ; Zufall & Rausher, ; Hoballah et al, ; Shimizu et al, ). Loss‐of‐function mutations often involve insertions or deletions in the coding region as a result of imprecise transposon exertion (Hoballah et al, ; Shimizu et al, ). In Petunia integrifolia , inactivating mutations in the ANTHOCYANIN2 ( AN2 ) gene which encodes a basic helix‐loop‐helix protein that directly activates DFR and converts dihydroflavonols into anthocyanins was found to be responsible for the absence of anthocyanin pigmentation, which converted the anthocyanin‐colored petals to white (Wijsman, ; Quattrocchio et al, ).…”

Cloning of the Flavonoid 3&apos;-Hydroxylase Gene of <i>Eustoma grandiflorum</i> (Raf.) Shinn. (<i>EgF3&apos;H</i>) and Complementation of an F3&apos;H-deficient Mutant of <i>Ipomoea nil</i> (L.) Roth. by Heterologous Expression of <i>EgF3&apos;H</i>

“…The progeny obtained by self-pollination of these cultivars were designated as ME, RV, and AK, respectively (Shimizu et al, 2011). Lisianthus plants were grown for gene cloning and RT-PCR analysis was performed as described previously (Hashimoto et al, 2004;Shimizu et al, 2011;Uddin et al, 2002). RNA was extracted from leaves as well as petals at six different floral developmental stages: 1) bud less than 20 mm long; 2) bud between 20 mm and 30 mm; 3) bud more than 30 mm long and tightly closed; 4) bud more than 30 mm long and loosely closed; 5) opening flower; and 6) fully opened flower.…”

“…The seeds of 'Mellow Lavender' and 'Royal Violet' were purchased from Takii Seed Co., Ltd. (Kyoto, Japan), and 'Asuka no Kurenai' was purchased from Sakata Seed Co., Ltd. (Yokohama, Japan). The progeny obtained by self-pollination of these cultivars were designated as ME, RV, and AK, respectively (Shimizu et al, 2011). Lisianthus plants were grown for gene cloning and RT-PCR analysis was performed as described previously (Hashimoto et al, 2004;Shimizu et al, 2011;Uddin et al, 2002).…”