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Molecular investigation of the progenitors, origin and domestication patterns of diploid Chinese old garden roses.

BACKGROUND AND AIMS: Chinese old garden roses are major contributors to the genetic development of modern roses. The RoKSN gene is associated with continuous flowering in roses and is proposed to have originated from Chinese wild roses. However, the wild roses that are implicated in the breeding of Chinese old garden roses and the origin of the RoKSN locus remain unidentified. We collected 25 of the most renowned and classic diploid Chinese old garden roses along with all related wild roses from East Asia. These roses were analysed with the aim of identifying the wild species that contributed to the genetic composition of Chinese old garden roses. In addition, we aimed to infer the geographical origin of the RoKSN gene and to develop a schematic overview of hybrid domestication of Chinese old garden roses. METHODS: We compared the haplotypes of internal transcribed spacers (nrITS), six nuclear single-copy genes and three chloroplast genes between Chinese old garden roses and wild roses. Additionally, we assessed genetic organization using 21 expressed sequence tag-simple sequence repeats to identify potential donor species that contributed to the emergence of these cultivars. Primers were designed for RoKSN to allow comparison of the gene across the entire distribution range of Rosa sect. Chinenses. KEY RESULTS: Our findings confirmed that the majority of rose cultivars are descendants of early hybridization events. Rosa chinensis var. spontanea, R. odorata var. gigantea and R. multiflora var. cathayensis were the primary donors for the 25 cultivar roses. Chinese old garden roses were categorized into four groups. Ten cultivars were hybrids between R. chinensis var. spontanea and R. multiflora var. cathayensis, thereby forming the 'Old Blush' group. Five cultivars were hybrids between 'Old Blush' and the R. kwangtungensis species complex, thereby forming the 'Slater's crimson' group. Six cultivars were hybrids between 'Old Blush' and R. odorata var. gigantea, thereby forming the 'Tea Rose' group, and three cultivars were hybrids that evolved from more than three donors. Moreover, we observed relatively close genetic proximity among Chinese old garden roses with an identical RoKSN-copia gene that is responsible for continuous flowering, which indicates a single origin for this retrotransposon-containing allele. Additionally, we determined that the haplotypes of the RoKSN-copia gene predominantly occurred in the Sichuan Basin region. In contrast, R. chinensis cultivated in the Ya'an region showed no markers of hybridization and displayed a genetic composition that was close to that of the wild species R. chinensis var. spontanea. This cultivar may represent the earliest mutated individual that bears the RoKSN-copia gene and may have served as a bridge from wild species to continuous-flowering old rose cultivars. CONCLUSIONS: The study provides crucial evidence that elucidates the origin of cultivated roses and lays the groundwork for further analysis of the breeding history of Chinese old garden roses using genomic data.

Domestication

RcAP2L-RcAS1 complex modulates petal number in roses by targeting RcAGL80 promoter.

Double flower, which is one of the most important characteristics of ornamental plants, is closely related to their ornamental and commercial value. The double-flower trait in rose was mainly due to the increase in petal number caused by stamen petalization. However, the mechanism regulating petal number is not clear. In this study, the Rosa chinensis "Zhaiye Tengben Yuejihua" × R. chinensis "Old Blush" population was used for QTL detection and NGS-based BSA analysis to identify candidate genes related to petal number. It was found that RcAP2L and RcAS1 were highly expressed in double-flower rose, while RcAGL80 was highly expressed in single-flower rose. Silencing RcAP2L and RcAS1 reduced the petal number by inhibiting homeotic conversion of stamens to petals, separately. However, silencing RcAGL80 increased the petal number by promoting homeotic conversion of stamens to petals. The results of Y2H and BiFC assays showed that RcAP2L interacted with RcAS1. The dual-luciferase assay showed that RcAP2L was bound to the promoter of RcAGL80 and suppressed RcAGL80 transcription. In total, we found a new function of AS1 in specifying flower organ identity, and a new pathway for regulating the number of petals by RcAS1, RcAGL80, and RcAP2L, which provides new information for elucidating the mechanism of the formation of double flower in rose.

Rosa

Study of the plate agglutination test with rose bengal antigen for the diagnosis of human brucellosis.

This study presents the results of a number of serological tests, in particular the plate agglutination test using acid antigen stained with rose bengal, in the examination of 440 human sera for brucellosis. In addition, the sera were examined for the presence of specific antibodies of various classes (IgM and IgG). These investigations showed the rose bengal plate agglutination test to be highly specific and sensitive as a rapid method for the diagnosis of human brucellosis.

Agglutination Tests

Relative efficacy of blood, urine, rectal swab, bone-marrow, and rose-spot cultures for recovery of Salmonella typhi in typhoid fever.

The recovery of Salmonella typhi from blood, rectal swab, urine, bone-marrow, and rose spots was compared in 62 patients with typhoid fever, most of whom had received some antibiotic therapy before presentation. S. typhi was isolated from culture of bone-marrow in 56 patients (90%); in contrast, S. typhi was recovered from blood in only 25 (40%), from stool in 23 (37%), and urine in 4 (7%). S. typhi was isolated from 24 (63%) of 38 patients who had rose-spot cultures. If culture sites had been limited to blood, stool, and urine, the bacteriological diagnosis would have been missed in 24 patients.

Bacteriuria

Vital staining of the corneal endothelium with rose bengal and alizarin red S.

A new technique for the staining of the corneal endothelium is presented, in which the actions of the vital stain, rose bengal, and the intercellular stain, alizarin red S, are combined. The technique is an improvement over other simple evaluative methods, because it defines both healthy and damaged cells, and thereby permits an investigator to accurately quantify endothelial cell damage.

Animals

Effect of NADP+ and its analogs on the Rose Bengal-sensitized photoinactivation of D-erythrulose reductase from beef liver.

Upon addition of NADP+, the rose bengal-sensitized photoinactivation of D-erythrulose reductase from beef liver is prevented to a remarkable extent. Adenosine 2',5'-diphosphate (2',5'-ADP) also has a protective effect, but to a lesser extent. On the other hand, 2'-AMP markedly enhances the photoinactivation. Other nucleotides which have no 2'-phosphoryl group, such as NAD+, 3'-AMP, 5'-AMP, ADP, and NMN, are ineffective. Further, only 2'-AMP derivatives (NADP+, 2',5'-ADP, and 2'-AMP) among these nucleotides were found to be potent competitive inhibitors of the enzyme with small Ki's (6--13 muM). Photooxidation of some methionine residues in the enzyme is prevented by the addition of NADP+ and accelerated in the presence of 2'-AMP. Photooxidation products(s) of 2'-AMP derivatives have no effect upon the enzymatic activity. Although NADP+ and 2'-AMP induce detectable conformational changes of the enzyme, the changes are not characteristic to the compounds. Based on these observations, we present a possible action mechanism of 2'-AMP derivatives on the photoinactivation of D-erythrulose reductase.

Adenosine Monophosphate

[Diagnostic value of agglutination reaction with buffered Brucella antigen stained with rose bengal].

A specific buffered antigen has been obtained, employing a method developed by the authors, stained with Bengal rose and intended for performing a fast agglutination reaction to confirm brucellosis. The antigen produces a clear and demonstrative agglutination reaction with positive sera. Practically, the test is readily carried out, and can be made a routine both in every serologic laboratory and for investigations under field conditions. The reaction produced with this antigen can determine dependably the epizootic status on a farm or in the herd. The diagnostic value of the antigen is essential, especially with swine. Besides, it shows a wide a diagnostic scope for brucellosis with all species of animals (97--98 per cent).

Agglutination Tests

[Interrelationship between glucoside-bound and free alcohols during ether oil formation in rose petals].

Some data on the dynamics of free and glucoside-bound monoterpenic and aromatic (beta-phenylethyl) ethers content and the changes in the beta-glucosidase activity in rose petals at different stages of the flower development and on the kinetics of enzymatic hydrolysis of these glucosides are presented. The phase specificity of beta-glucosidase coinciding with the maximal accumulation of glucoside-bound and free alcohols is revealed. The data obtained suggest that the formation of glucosides may precede the accumulation of corresponding free alcohols of terpenic and aromatic origin.

Alcohols

[Biosynthesis of beta-phenylethanol in rose petals].

Biosynthesis of beta-phenylethanol is studied in a cell-free rose petals extract. This compound found to be synthesized via two intermediates: either trans-cinnamic acid, or phenylacetaldehyde. Subsequently beta-phenylethanol is glucosodized into its transport form, beta-phenylethanol- beta-D-glucoside. A scheme of beta-phenylethanol and its beta-D-glucoside is given.

Acetaldehyde