Genetic Study of Olive Plants

Genetic Study of Olive Plants Introduction Oleaceae family comprises 24 genus which consists of 600 species and its members can be found all over the world. One of the first cultivated agricultural tree crops within Oleaceae family is the olive Olea europaea L which is usually used for production of table olives and edible oil. Olive is the native plant in coastal regions of Mediterranean and there are more than 86 varieties of Europea species found in Anatolia of Turkey. Turkey is known to be one of the largest producers of olive oil and the first producer of black table olive among all the countries in the world. Majority of the black table olive that produced by Turkey is the cultivar of Gemlik which represents 80% of the production. Therefore, many researches in Turkey are mainly focused on molecular and classical breeding program for Gemlik cultivar due to its economical importance. It is essential to have the genetic studies in olive plants in order to understand the genetic mechanisms and hence to improve the quality and quantity of products. Expressed sequence tags (ESTs) which are obtained from the single-pass complementary DNA (cDNA) sequencing can provide genetic information of an organism. Since EST is generated with gene that expressed at certain stage or tissue of organism, the information on mature transcripts for coding region of genome can be shown by EST. Hence, EST databases are useful for functional studies, gene and marker discovery, and gene mapping. Currently, EST libraries for more than 40 plant species are established for providing information that is important for functional genomics studies in which the putative functions of genes can be deduced by searching for homologies with known genes. Before this study, there were just around one thousand EST sequences which were available in GenBank database on February 2009. The EST studies for olives are obviously insufficient and lacking of sequence information may limit the genetic studies in olive plants. In this paper, two separate cDNA libraries which were constructed from young leaves and olive fruits for olive cultivar Gemlik were used to establish a rich EST collection. Clones which were obtained from the cDNA libraries were sequenced to generate ESTs. These sequences were assembled and clustered by using specific software and they were then deposited in GenBank database. Annotation of the generated EST sequences was performed using BLAST and BLAST2GO. Method of preparing Expressed Sequence Tag (EST) database The research material used in this study was the olive (O. europea) cultivar Gemlik (G 20/1). The methodology that involved in preparing EST database for Olea europaea was described by Ozgenturk et al. (2010). Before constructing cDNA library, total RNA was isolated from young leaves and unripe olive fruits by using the RNA extraction kit. In this paper, RNeasy Plant Miniprep kit was used for total RNA extraction. Total RNA was treated with the Oligotex mRNA Mini Kit to obtain purified mRNA. The isolated mRNAs were collected and utilized for constructing two separate cDNA libraries. Since RNA molecules are very unstable and difficult to amplify, it is important to convert mRNAs into cDNAs which are more stable in order to analyse the information carried by mRNA molecules (Mullinax Sorge, 2003; Tovey, 2011). The cDNA libraries can be constructed with the isolated mRNAs by using a cDNA library construction kit and the CloneMiner cDNA Library Construction Kit was used in this paper. Briefly, a double-stranded cDNA (ds cDNA) was first synthesised from mRNA by using primers provided in the kit. For the first strand synthesis, biotin-attB2-Oligo(dT) primer bound to the poly(A) tail of mRNA and the mRNA was used as the template for reverse transcription by SuperScriptTM ΙΙ Reverse Transcriptase. The newly synthesised single-stranded cDNA (ss cDNA) served as the template for second strand synthesis by Escherichia coli DNA Polymerase ÃŽâ„¢. attB1 sequence was then incorporated to the 5’ end of ds cDNA by attB1 Adapter. The synthesized ds cDNA was ligated into pDONR222 vector and transformed into competent E. coli strain DH5. The two cDNA libraries were plated onto LB-kanamycin agar plates. Individual colonies formed on the plates were picked into 384-well plates which containing SOB medium and incubated overnight. The cDNA libraries were then stored at -80 °C after adding with glycerol (10% v/v). Complementary DNA (cDNA) clones were randomly selected for isolating plasmid DNA with alkaline lysis method. The isolated DNA was digested and insert size was identified by performing 1% agarose gel electrophoresis. In this paper, 3840 clones were randomly selected and amplified by polymerase chain reaction (PCR) using M13 universal primers. The ABI 3730 capillary sequencer was used for automated sequencing of cDNA in order to generate ESTs. Results Two separate cDNA libraries were established with mRNA extracted from young leaves and fruits respectively. The average insert sizes for the leaf and fruit cDNA libraries were respectively 1.6 kb and 1.1 kb. The leaf cDNA library consisted of 2.4 Ãâ€" 106 clones and 2304 clones were selected for sequencing. Among 2.2 Ãâ€" 105 clones within olive fruit cDNA library, 1536 clones were sequenced. Therefore, altogether 3840 EST sequences were generated from cloned cDNA sequencing. In order to obtain high quality EST sequences, raw EST sequences were processed by using Phred software in which low-quality bases, vector, and adapter were removed. After processing, 106 low quality EST sequences were removed and the remaining 3734 ESTs were kept for contig assembly. Contig Assembly Program 3 (CAP3) was used to assemble leaf and fruit EST sequences into contigs. 205 contigs were generated from the assembling of 2228 leaf ESTs and 69 contigs were generated from 1506 fruit EST sequences assembly. The total numbers of contig and singleton that were established from these two libraries were 274 and 2478 respectively. Altogether 3734 ESTs and 249 high quality contigs were deposited in GenBank under the accession number GO242703 to GO246436 and EZ421546 to EZ421794 respectively. In order to designate annotation of the generated EST sequences, BLAST was performed on the National Center for Biotechnology Information (NCBI) web server for searching gene homology. Six hundred thirty five EST’s unique gene sequences were found to have more than 80% homology with sequences of known function in the other species. Two thousand twenty four EST sequences have less than 80% homology to the expressed proteins, hypothetical proteins, putative uncharacterized proteins and unknown proteins in database. There are 1339 EST sequences that shown no homology with the sequences that exist in GenBank and about 96.9% of EST sequences that established in this study are different from that in the existing olive sequences database in NCBI. Therefore, these EST sequences can be considered as novel genes in O. europea. Besides BLAST analysis, gene ontology (GO) annotation was performed for the contig sequences generated from the ESTs by using Blast2GO. GO terms were distributed among the biological process, molecular function and cellular component categories.

Management and University Simmons Case Essay

In time of economic crisis, companies are looking for innovative methods to improve production and to meet the needs of a diverse workforce in order to improve or maintain the organization’s profit margin in a stress global economy. Indeed, companies are extremely concerned about their future, as well as, preventing closure of their establishment. SimmonsCompany is no exception. However, how does a major company make those changes when tradition is the foundation of their organization and the economic status of their company is in jeopardize of folding, if the right decision is not made? The need to allocation funds for the training and development of its and employees when major debts are owed to the company; thus, threatening bankruptcy is a problem facing Simmons Company. Accordingly, there is certainly a need to change the culture and structure of the organization, if it wants to survive a depressed economy, while other organizations are advancing. In so doing, the company is considering the Great Game of Life (GGOL) whose underlying purpose is to change the dynamics of an organization in order to achieve maximum satisfaction for both the customers and employees. If customers and employees are satisfied, they are loyal, cooperative and creative, thus creating a culture conducive to the workforce. In order to establish such an environment, it is necessary to empower its workers by changing the attitude of each individual employed by Simpson Company. The program will be used to improve morality and empower the lower level of the company in performing their respective jobs in order to improve the outcome of the company. Changes are often difficult to make and, are usually resisted from the top because control of the company is centralized. The result is a poor culture and dictatorial practices. Immediate changes are normally discouraged by those in control, but gradual changes are encouraged, if done overtime or gradually phased into the daily operation of the company. Management must recognize that if changes are not made, their position might be null and void for not doing so. The question is whether changing the culture of the organization is in its best interest? Indeed, some changes must be made considering the state of fair of the company. There are several outstanding debts owed to SimmonsCompany and the debtors are threatening bankruptcy. Additionally, a major supplier has caused havoc for the production department. The need to change the culture and the way it operates is more apparent than ever before. To stay the same, means eventual failure for the company. GGOL is a great opportunity to enhance or change its culture and improve its staff’s level of expectation through effective management in order to meet the needs of today’s demanding economy. As previously mentioned, the company is already experiencing a decline in its profits margin because of its accounts receivables from customers and their major supplier has an item in the form, emitting afoul odor, causing the company to compromise production schedules and posed a serious threat to its profit margin. It would certainly influence my decision to implement GGOL at Simmons. The video was both inspirational and informative. Since diversity is a major concern for most global companies, this is not the case with Simmons. That is, it was prevalent at Simmons, as well as, their enthusiasm in working with each other at the company and for the company. Their positive attitude resonated throughout the video. It is evident from the video that they work collectively and are eager to assist others in different department, when asked to do so. Coming to work seems second nature for them. They are able to share with the upper management areas of concerns and questions, and as such, not prohibited from doing so, which was not the case in the past. In short, they are empowered to take on task without being micro-managed. Now, it is a shared vision by the top, middle, and lower levels of the company with one mission in mind to do what is in the best interest of company. The company should use the top-down and bottom-up design in implementing the GGOL program. However, the company should employ a professional evaluation team to determine which plant should be the catalyst to receive the program. After selecting the plant, the program should start at the top level of the plant because they are responsible for the day-to-day operation of the plant and if they buy into the program other levels are more willing to accept it. The top-level can assist in delivering the GGOL program to the other employees. The next level should be the middle level because they are responsible for inspiring the workers to work collectively towards a common goal, i.e., what is in the best interest of the plant. Needless to say, these individual are very reluctant to change, but must be done gradual due to their commitment to the company, embedded old core values, and long history with the company. When the worker realizes that the change is positive, their outlook improves and this attitude has a snowball effect. That is, one worker at a time starts to embrace middle management’s vision for the company. Now, the workers are willing to trust middle manager because they are open for suggestions and are encouraging empowerment of workers, unlike the past, when the decision from the top was purely totalitarianism with no questions asked or else be fired. Finally, the workers should be the last to receive the program. The middle-level can introduce the program. In so doing, it encourages team work, which will ultimately change the culture of the plant. Changes are needed and welcomed, if it benefits both the employer and employees. If this company continues to operate in a vacuum, the destiny of the company is almost sudden death with uncollectible debts arising and plant processing being restricted because of the foul odor. Emotions were running high, each level of the organization realizing the potential of closure of the company, if it did not change the culture of the organization. The old way of handling concerns and questions is pseudo at best. The company would continue to experience apathy at the workplace. Thus, the $7 million dollar investment for the training and development of personnel is an extremely prudent investment for the company. References Kotter, J. P. (1994). Leading Changes. Boston, MA: Harvard Business School Press. Leading Change at Simmons (A)http://gcumedia.com/digital-resources/harvard-business-school-press/2007/change-at-simmons-part-a_1e.php Leading Change at Simmons (B)http://gcumedia.com/digital-resources/harvard-business-school-press/2007/change-at-simmons-part-b_1e.php

The Righteous Deceit of Helen Turrell

Helen Turrell is portrayed at first glance as an independent caretaker of her nephew, Michael Turell in the short story, â€Å"The Gardener† by Rudyard Kipling. However, upon reading between the lines of this character's story, her facade of honesty is peeled away. In it's place is a of lies exposed by the story's namesake, the Gardener. Her experience with the public is through a veil of her lie. Instead of going to France to cure her lung trouble as the public thought, Helen was actually there bearing Michael into the world. Besides this secret, Helen's honesty is a well-known trait to the general public.To her son, however, her honesty and lies are a strain on their relationship. By ten years old he discovers that he is an illegitimate child. When Michael unexpectedly dies fighting in World War I, Helen meets Mrs. Scarsworth. They meet while traveling to visit Michael's grave, and Mrs. Scarsworth prompts the first introspection of Helen's lie. The Gardener at the graveyard confirms the illegitimate son to the reader. The story seems to be told from Helen's version of the happenings, and the public's gossip about it. Outwardly, the public's gossip defines Helen's appearance outside of her deceit.Rudyard Kipling builds on his theme of the effects of deceit buy developing the character Helen Turrell into an innocent, selfless, yet deceitful person through her twisted interactions with the public in general; her unfulfilled relationship with Michael Turrell; and her short, strained relationship with Mrs. Scarsworth. Helen's relationship with the public was a deceitful one, but both parties gained from the relationship. The beginning paragraphs of the story aren't quite from the authors omniscient narrative. The sentences are short and information is sparse and sometimes absent, as though it was gathered through gossip.From this it can be concluded that the public gossip is writing this part of the story. The opening paragraph explains that Helen Turrell i s sister to recently deceased George Turrell; when he died he left a scandal in his wake of an illegitimate baby in India. Helen was suffering health issues and recovering in France, but she returns back to her home in Hampshire with her nephew Michael. As far as her relationship with the public, â€Å"All these details were public property, for Helen was as open as the day,† and â€Å"scandals are only increased by hushing them up† (Kipling).Her honest reputation shows how well the Helen kept her secret from the public. The gossip on Helen seemed to sometimes focus on how much of an honest person she was, and this can be interpreted in two ways. Her honesty made the public respect her relationship with her nephew, even though she reveals that she lets him call her â€Å"mummy† at bedtime. No foul play was suspected. However, focusing on how honest someone is begs for attention to the fallacious human condition, and this is a subtle hint to the other side of Hel en's facade. This may have eaten away at her, causing some of the distress in her relationship with her son.Helen Turrell has a unique relationship with her son that significantly defines her character into one of innocence and goodwill despite her facade. When her son, Michael, is around 10 years old, he realizes that his â€Å"civil status [is] not quite regular,† and then he proceeds to â€Å"[break] down her stammered defences† (Kipling). The question of Michael's legitimacy will obviously be a sore point for Helen that she isn't interested in talking about much. She has been hiding it her entire life, and that is not a small feat, for it is her own son!This, combined with allowing Michael to call her â€Å"Mummy† at bedtime shows that she still considers him her son, with all the attachment and emotion that comes with it. After hiding it for so long, Helen doesn't have anything other than a tentative response to Michael's attacks. Her tentative response ref lects that Helen does not dwell on her deceit. She understands that deceit is morally incorrect, and she is embarrassed by its evil nature. Because Helen is somewhat foreign to her lie, her innocence is preserved through the amoral facade. Her lie is so her son can have a better life; it is selfless, and therefore Helen is arguably moral.This isn't to say that Helen regrets her predicament as Helen's meetings with the flustered Mrs. Scarsworth reveal. Helen's relationship with Mrs. Scarsworth shows that Helen almost doesn't realize the extent of her deception, and her deception is further shown as good compared to Mrs. Scarsworth's pathological lying. Helen meets Mrs. Scarsworth through her journey to visit the grave of the now deceased Michael. Mrs. Scarsworth is visiting the graveyards under the premise of seeing commissions for friends who would be comforted knowing someone made the trip. She ends up staying in the same hotel as Helen. In the middle of he night, Mrs. Scarsworth d isturbs Helen to confess that one of her commissions was her love. Helen â€Å"desperately† asks, â€Å"But why do you tell me? †(Kipling) Helen's desperation is an obvious response to being disturbed so late at night. It is obviously annoying to be awoken in order to tend to someone's moral dilemma, but Helen is paying more than flustered attention to the problem of Mrs. Scarsworth. Helen's lie is close to revealing herself at the question of why Mrs. Scarsworth would reveal to her. Helen fears that Mrs. Scarsworth feels an inherent kinship with Helen and the night time intruder might guess at Helen's deception.That is why Helen truly feels desperation towards Mrs. Scarsworth. Helen shouldn't be worried though, for Mrs. Scarsworth confesses of â€Å"always lying† for about â€Å"six years†(Kipling) of deception total. This pathological lying is a circumstance of evil that serves as an antithesis of Helen's situation. Mrs. Scarsworth is visibly and audibl y troubled by her lying, whereas Helen was only troubled by the necessary strain it put on her relationship with her son. Helen has lived with the deception for so long without worrying as much about it that it doesn't matter so much to become the crippling and defining trait that it is in Mrs.Scarsworth's life. Through Helen's emotions in her relationship with Mrs. Scarsworth, it is really revealed that Michael is her son and not her nephew. Here there is no mention of public gossip, but perhaps Helen can now embrace her innocence and confess now that Michael has passed away. Helen's character is developed as such through her relationships with the other characters in this short story. Her development is especially shown in her unique interactions with each different character because of her deception. But in the end, her deception is an act of selflessness and love, and cleans Helen's moral slate.As a character, Helen's selfless innocence is finally proved by the appearance of the gardener. The gardener's â€Å"endless compassion† â€Å"Unending love† in his eyes when his omniscience shows Helen where her â€Å"son†(Kipling) was redeems her. The Gardener's traits match Christian Christ himself, and in showing Helen where her son is, forgives her. The story ends here, but if it was told in reference to what public gossip knows, Helen must have confessed after her trip to the graveyard. Thus, Helen finally opens up and moves on with her life past her righteous deceit.

Childhood Obesity A Growing Problem - 1378 Words

Did you know that obesity has become such a ‘growing’ quandary that for the first time ever today’s children are expected to have a life expectancy that is shorter than their parents (Gance-Cleveland, Gilbert, Kopanos, Gilbert, 2009, p. 72)? Obesity in children and adolescents has become a worldwide epidemic, increasing every year. In fact, childhood obesity in the United States (US) has increased to 17%, which has nearly tripled the prevalence of obesity in the last three decades (Center for Disease Control and Prevention [CDC], 2012). Childhood obesity is a significant issue, with many of its adverse health consequences often continuing into adulthood. The adverse affects of obesity are associated with hypertension, type II Diabetes Mellitus, cardiovascular disease, cancer, and numerous other comorbidities (Gance-Cleveland, et al., 2009, p. 72). According to Cornette (2008), there are also â€Å"non-physical consequences of childhood overweight such as depression, social isolation, discrimination, and poor self-esteem, self-image, academic performance, and professional performance† (p. 136). These comorbidities, inevitably, lead to an exponential growth in obesity-related health care costs. As healthcare professionals, we need to address the issue of childhood obesity in order to avoid further damage to those affected, decrease obesity in our youth, as well as decrease the healthcare costs that coincide with obesity. This paper focuses on the multitude of factorsShow MoreRelatedChildhood Obesity. Child hood Obesity Is A Growing Problem1085 Words   |  5 PagesChildhood Obesity Childhood obesity is a growing problem in the United States. Childhood obesity is defined as a child being at or above the 95th percentile for the child’s age and sex (Dietz, 1998). 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