Ampelographic features of biotypes of ‘Saperavi’ grape variety

Целью работы является установление отличий выделенных биотипов сорта Саперави по основным ампелографическим признакам, а также обсуждение использования термина «биотип» в виноградарстве и возможности практического применения биотипов винограда. В результате исследования насаждений сорта Саперави выделено 4 биотипа, включая контрольный биотип. Биотип I: гроздь ветвистая, коническая, рыхлая, средней величины, длина грозди 13-15 см, масса грозди 180-220 г, ягода мелкая и округлая. Биотип II: гроздь ветвистая, коническая, рыхлая, длина грозди 16-18 см, масса грозди 270-320 г, ягода средней величины и продолговатая. Биотип III: гроздь ветвистая, ширококоническая, большая, длина грозди 19-21 см, масса грозди 500-600 г, ягода крупная и овальная. Биотип IV (контроль): гроздь ветвистая, коническая, рыхлая, длина грозди 17-19 см, масса грозди 330-450 г, ягода средней величины и овальной формы. Среди изученных биотипов наиболее перспективным является биотип III по признакам величины, плотности и массы грозди, размера ягоды и выхода сусла. «Биотип» - это термин, употребляемый для альтернативного обозначения клона, группы клонов или сорта винограда, используемого в определенном регионе. Концепция биотипа находит свое применение в экспериментальных исследованиях и в клоновом отборе при необходимости подчеркнуть уровень изменчивости более высокой, чем у сорта или клона. Полученные результаты могут использоваться при возделывании сорта Саперави в виноградарских хозяйствах, а также в виноделии. Determination of the differences between the selected biotypes of ‘Saperavi’ variety according to the main ampelographic traits, as well as discussing the use of the term “biotype” in viticulture and the possibility of practical application of grape biotypes are the aims of the work. As a result of the study of ‘Saperavi’ variety vineyards, 4 biotypes were identified, including the control one. Biotype I: the bunch is branched, conical, loose, of a medium size, the bunch length is 13-15 cm, the bunch weight is 180-220 g, the berry is small and round. Biotype II: the bunch is branched, conical and loose, the bunch length is 16-18 cm, the bunch weight is 270-320 g; the berry is medium-sized and oblong. Biotype III: the bunch is branched, broad-conical and large, the bunch length is 19-21 cm, the bunch weight is 500-600 g; the berry is large and oval. Biotype IV (control): the bunch is branched, conical, loose, the bunch length is 17-19 cm, the bunch weight is 330-450 g; the berry is of medium size and oval. Biotype III was the most promising one among the studied biotypes according to the size, density and bunch weight, the berry size and the yield of must. “Biotype” is a term used to alternatively denote a clone, group of clones or grape variety used in a certain region. The concept of a biotype finds its application in experimental studies and in clone selection, if necessary to emphasize the level of variability higher than that of a variety or clone. The results obtained can be used in the cultivation of ‘Saperavi’ variety in vineyards, as well as in winemaking.

Artificial Neural Networks and Linear Regression Reduce Sample Intensity to Predict the Commercial Volume of Eucalyptus Clones

Equations to predict Eucalyptus timber volume are continuously updated, but most of them cannot be used for certain locations. Thus, equations of similar strata are applied to clonal plantations where trees cannot be felled to fit volumetric models. The objective of this study was to use linear regression and artificial neural networks (ANN) to reduce the number of trees sampled while maintaining the accuracy of commercial volume predictions with bark up to 4 cm in diameter at the top (v) of Eucalyptus clones. Two methods were evaluated in two scenarios: (a) regression model fit and ANN training with 80% of the data (533 trees) and per clone group with 80% of the trees in each group; and (b) model fit and ANN training with trees of only one clone group at ages two and three, with sample intensities of six, five, four, three, two, and one tree per diameter class. The real and predicted v averages did not differ in sample intensities from six to two trees per diameter class with different methods. The frequency distribution of individuals by volume class by the two methods (regression and ANN) compared to the real values were similar in scenarios (a) and (b) by the Kolmogorov–Smirnov test (p-value > 0.01). The application of ANN was more effective for total data analysis with non-linear behavior, without sampled environment stratification. The Prodan model also generates estimates with accuracy, and, among the regression models, is the best fit to the data. The volume with bark up to 4 cm in diameter at the top of Eucalyptus clones can be predicted with at least three trees per diameter class with regression (root mean square error in percentage, RMSE = 12.32%), and at least four trees per class with ANN (RMSE = 11.73%).

Dynamics and Characteristics of BCR-ABL1 Multiple Mutations in Tyrosine Kinase Inhibitor Resistant CML

Abstract 1677 Background: BCR-ABL1 kinase domain (KD) point mutation causes resistance to tyrosine kinase inhibitors (TKI) in CML patients through impaired binding of TKI to the target site. One of the characteristics of patients with BCR-ABL1 kinase domain point mutations is the fact that some patients have multiple mutations. However there have not been many studies showing that data about clinical relevance or dynamics of multiple mutation during CML treatment. Methods: Since 2002, 414 CML patients were screened for mutation analysis due to sign of resistance to TKI including imatinib, nilotinib, dasatinib, bosutinib, radotinib or ponatinib at Seoul St Mary's Hospital using direct sequencing and ASO-PCR. Among them, 31 patients showed multiple mutations. We analyzed serial samples from the 31 patients using subcloning and sequencing to investigate whether the multiple mutations are on same clone (defined as compound clone), separated clones (defined as multiple clone) or co-existent clones (defined as mixed clone) and characterize its clinical relevance and dynamics. Results: Status of the patients with multiple mutations is shown in Table 1. In order to investigate whether the multiple mutations are on same clone or on separated clone, we cloned serial samples from the 31 patients. Cloning of cDNA region corresponding to BCR-ABL1 KD into plasmid was performed and followed by transformation into competent cells, colony formation, plasmid preparation of 20 colonies from each sample, and then direct sequencing. Multiple mutations of 65% patients (20 out of 31) existed compound mutation which means the individual mutant types are located on the same BCR-ABL1 molecule. In addition of major mutation types which were detectable in direct sequencing analysis, all the patients showed to have minor types of mutations which were found only through BCR-ABL1 KD cloning and subsequent colony sequencing. To make sure that this minor mutation types were not caused by sequencing error, we also analyzed of 3 patients who showed TKI resistance, but had no BCR-ABL1 mutation. In addition, samples from 3 normal persons were analyzed with the same method. The frequency of appearance of the minor types of point mutation was reduced in the patient group who showed TKI resistance, but had no BCR-ABL1 mutation, and then dramatically decreased in the normal person group, indicating that BCR-ABL1 gene in patients with point mutation are relatively unstable. Among 20 patients with compound mutation, 9 patients were available for serial timepoint samples under same TKI therapy. In all nine patients (100%), portion of compound clone was increased as treatment went on. With a median follow-up 53.3 months (range, 0–113.2 months), of 31 patients with multiple mutation, 7 patients remained alive; 4 of 11 (36%) in the multiple clone group vs 3 of 20 (15%) in the mixed clone group (P = 0.066). Conclusion: Analysis of serial samples from a same patient provided evidence of dynamic change of portion of compound mutation. In most case, portion of the clone containing compound mutation was increased as treatment went on, indicating the clone harboring compound mutation can take survival advantage over TKI treatment in comparison of the clone containing individual type of mutation. In addition, some patients showed change in individual mutation type comprising multiple mutations as treatment went on. Patients with compound clone showed poor outcomes compared with multiple clone group in our cohort, further investigation on a large patient cohort will be needed. Updated data with longer follow-up duration will be presented in the meeting. Disclosures: No relevant conflicts of interest to declare.

Survival Analysis of Additional Chromosomal Aberrations in Philadelphia Chromosome Positive and Negative Cells: A Single Institutional

Abstract 2523 Background: Chronic myeloid leukemia (CML) is a clonal stem cell malignancy associated with the Philadelphia chromosome, t(9;22)(q34q31)/BCR-ABL gene fusion. Additional cytogenetic abnormalities have been known to emerge in Philadelphia (Ph) positive cells (additional Ph+ clones) or Ph negative cells (Ph- clones) at diagnosis, or during or post tyrosine kinase inhibitor (TKI) therapy. Many studies have elucidated that the presence of some Ph+ clones were frequently associated with disease progression, while presence of Ph- clones were unrelated to disease outcome. A majority of studies have shown a waxing and waning of Ph- clonality in response to therapy. However, there remains no solid data regarding overall survival comparing Ph+ and Ph- clones over a long term period. This study focuses on the observation of clonal evolution in various phases of CML, and the relationship to overall survival, disease resistance, kinase domain mutation (KDM) and progression to accelerated or blast phase in patients with CML. Materials and Methods: Data from 318 patients who were diagnosed with CML was retrieved from Moffitt Cancer Center during January 1990-December 2010. Patients are divided into three groups based on the presence of additional ph+ clone (A), ph- clone (B) and absence of both (C). Clinicopathologic results including initial diagnosis date, nature and frequency of clone, copy of clone at karyotyping, disease status in response to treatment and overall outcome were documented and statistically analyzed. All patients with Ph- and Ph+ clones have been treated with TKIs since 2001. Median overall survival and median disease progression survival were compared between conal ph- and clonal ph+ group by the log-rank test. Survival curves were generated using the Kaplan–Meier method. All reported P values are two-sided. Cox Regression (multivariate analysis) was also performed for time to progression. Results: Of 318 (average age 57, range: 19 to 89, M:F=1:1), 17 carried ph- clones (5.3%) and 41 showed additional ph+ clones (12.9%) and the rest (258) lacked additional ph+ or ph- clones. Additional clonal cytogenetic aberrations were random with the most frequent occurrence of trisomy 8 (7 of 17 ph- and 14 of 41 ph+ clones, respectively) and isochromosome 17q (4 in ph+ clone only). There is a higher rate of transformation to accelerated/blast phase in Ph+ additional clones (36.6%,15 of 41) as compared to Ph- additional clone (17.6%, 3 of 17) and Ph+ without additional clones(11.1%, 29 of 260) (p=0.015). The overall median survival is shorter in patients with ph+ clones (133.4 months) than in those with ph- clones (172 months) (p<0.005). KDM were observed in 27 of 110 tested patients (24.5%), and many of them fell into the Ph+ clone group (10 of 22 tested) and fewer into ph- clone group (3 of 10 tested) (p<0.005). Statistical analysis proved that there is a higher rate of transformation to accelerated/blast phase in the presence of a KDM (p<0.005). Of note, most of those with ph- clones had only one episode (59%) with 41% having more than one occurrence and lasting from 6–58 months (average 21.1 months), while in those 41 patients with Ph+ clone; a subset (18 of 41, 43.9%) showed a long-lasting ph+ clone over months (ranging from 2 months to 70 months, average 14.5 months) and the remaining only occurred one occasion (23 of 41, 56.1%). Multivariate analysis for disease progression demonstrated statistical significance with regards to both the ph+ clone and kinase mutation categories (p=0.003 and 0.034). Conclusion: Most Ph+ or Ph- clones are occasionally observed during the long course of CML, with a minor subset of clones showing persistency, especially in Ph- ones. The presence of an additional Ph+ clone is correlated with disease progression, drug resistance and shorter overall survival in comparison to a Ph- clone. A relatively long overall survival is observed in patients with ph- clones. Ph- clones were often identified during disease remission regardless of frequency of clonal copy or clonal persistence. The outcome of patients with concurrent Ph+ and Ph- clones was dictated by the Ph+ clone. Disclosures: No relevant conflicts of interest to declare.