Factors Affecting Organogenesis in Plant Tissue Culture

Factors Affecting Organogenesis in Plant Tissue Culture Organogenesis is the process of forming a specific organ from non-specific mass of meristem or parenchyma cell known as callus. In this experiment, auxin and cytokinin are used to determine the effect of PGR, auxin and cytokinin on the organogenesis in carrots and petunia leaves, in which auxin hormones responsible for growth of roots, phototropism and gravitropism, while, cytokinin helps in inducing the growth of shoots and regulates auxin action. Theoretically, the presence of high cytokinin and low auxin in the media, the shoots will form, meanwhile, in low level of cytokinin and high level of auxin in the media, roots will form. Meanwhile, in the presence of high concentration of both cytokinin and auxin, callus formation will be induced. However, in this experiment, production of the shoots and roots are not compatible with the theory due to improper ratio of auxin and cytokinin and the high toxicity of synthetic cytokinin, kinetin. Introduction Plant tissue culture (PTC) is the techniques used to grow plant from any of the plant segment, tissues or cell in a contaminated free environment media such as MS media (Singh Kumar, 2009). PTC techniques is important plant biotechnology aspect in which it facilitates the production of genetically modified plants and induced rapid multiplication of difficult-to-propagate plant species. Besides that, the ability to produce totipotent plant cell using PTC techniques has significant impact on crop improvement via genetic engineering (Radzan, 2003). There are two different processes, which involve explant differentiation and growth in PTC, which are organogenesis growth and adventititous roots or shoots growth directly from the explants. Organogenesis is the process of forming a specific organ from non-specific mass of meristem or parenchyma cell known as callus. Meanwhile, for formation of adventitious roots or shoots means the roots or shoots structure arise from the explants that have been excised (Pernisova et al., 2009). This situation does not usually happen if the plant sample are cultured in a medium with the same ratio of auxin and cytokinin. The presence of plant growth regulators (PGR) such as auxin, cytokinin, gibberellins, abscisic acid and ethylene has significant impact in the process of plant growth and differentiation. Gibberellins responsible for growth, seed germination and promote fruits growth. As for ethylene helps in controlling of fruit ripening as well as controlling cell division and cell elongation. Meanwhile, abscisic acid act on seed maturation and give the ability to the seeds to response during stress in undergoing dormancy period (Davies, 2010). However, in this experiment only, auxin and cytokinin are involve, in which auxin hormones responsible for growth of roots, phototropism and gravitropism, while, cytokinin helps in inducing the growth of shoots and regulates auxin action (Davies, 2010). In this experiment, NAA will be used as synthetic auxin and kinetin and BAP as synthetic cytokinin. This experiment was conducted in order to determine the effect of PGR, auxin and cytokinin on the organogenesis in carrots and petunia leaves. Materials and methods Plant materials For petunia leaves samples, the leaves were provided by lab technician in Monash University Sunway Campus. Firstly, 18 petunia leaves were collected from the petunia leaves petri dish. Then, the petunia leaves were soaked using 10% (v/v) of sodium hypochlorite for not more than 5 minutes. The leaves were then rinsed using sterile water in a laminar flow cabinet. Then, 2 petunia leaves were added in each petri dish with different ratios of NAA:BAP and NAA:Kinetin. The same methods were implied to the carrot. However, the carrot sample was cut into 27 pieces about 0.5cm thick on the surface of sterile ceramic tile. After soaking the carrot samples in sodium hypochlorite and rinsed for three times, 3 carrot pieces was transferred into petri dish with different ratios of NAA:BAP and NAA:Kinetin. The result recorded based on table 1 and table 2. Tissue culture media MS media supplemented with combination of high cytokinin to low auxin were prepared by the lab technician as follows: NAA:Kinetin ratios (2:0, 0.5:1, 1:0.5, and 0:2) and NAA:BAP ratios (2:0, 0.5:1, 1:0.5, and 0:2). Additional of two control plate with MS media that supplemented with same ratio of auxin and cytokinin were also prepared by the lab technician. Culture condition The transferred petunia leaves and carrot pieces in the 18 petri dishes were incubated for 28days in 25ÂÂÂ ±2ÂÂÂ °C temperature, and photoperiod of 16hours in light 8hours in dark as provided in plant culturing room in Monash University Sunway Campus. In addition, for every 4 days, the tissue cultured was checked to whether there are presents of contamination and to transfer the samples into new MS plates. Results and discussion Effect on carrot slices Based on the result obtained in table 1, it shows that for carrot samples, the formation of callus can be seen all of petri dish with various ratios of either BAP:NAA or Kinetin:NAA. Theoretically, the formation of callus is due to the high concentration of both the cytokinin and auxin in a growth media (Duncan et al., 1985). Since all the plates are containing callus, it can be deduced that the growth of callus can be formed by excision of the plant cell. Meanwhile, for formation of roots, only carrot samples from MS media supplemented with BAP:NAA with ratio of 0.0:2.0 which also shows formation of roots. This is because, according to the theory, in the presence of high cytokinin and low auxin in the media, the shoots will form, meanwhile, in low level of cytokinin and high level of auxin in the media, roots will form. However, in the presence of high concentration of both cytokinin and auxin, callus formation will be induced (Chawla, 2002). However, it can be seen that there is no production of shoots in other plates except in MS media containing 0.0BAP:2.0NAA. This may occur due to unsuitable ratios of cytokinin and auxin in MS media that may not favorable for carrot species to induce formation of shoots. Effect on petunia leaves By referring to table 2, it shows that petunia leaves sample that shows formation of callus, roots and shoots is the MS media supplemented with BAP:NAA with ratio of 0.5:1.0 and the control plate. This occur due to suitable ratio of cytokinin and auxin suplemeted in the MS media. Furthermore, only MS media supplemented with BAP:NAA with ratio of 0.0:2.0 shows the growth of shoots excluding the MS media supplemented with BAP:NAA with ratio of 0.5:1.0 and the control plate. Theoretically, the productions of shoots are triggered by high concentration of cytokinin and low concentration of auxin. However, in the ratio of 0.0BAP:2.0NAA, it is predicted that formation of roots will be induced, instead, shoots are forming. This occur due to presence of zeatin in petunia leaves. Zeatin is a natural cytokinin presence in the most green leaves that undergoing senescence (Singh et al., 1992). The presence of cytokinin in the media may mask the reaction of auxin hormone on the explants presence i n the media (Nakagawa et al., 2006). Therefore, the production shoots are triggered instead of roots. Based on the result in table 2, the production of roots and shoots are low due to due to the excessive amount of alcohol used during the sterilization process. Besides that, shoots and roots only produce in MS media supplemented with BAP:NAA and not in MS plate with Kinetin:NAA. This may occur due to the toxicity of the synthetic kinetin, that are not suitable for the growth of either shoots or roots on petunia leaves. There were also presence of contamination in the cultured plate of carrot sample, which involve MS media supplemented with BAP:NAA with ratio of 2.0:0.0 and MS media supplemented with Kinetin:NAA with ratio of 2.0:0.0. Meanwhile, for petunia leaves sample, the cultured plate that have been contaminated by fungal infection are MS media supplemented with BAP:NAA with ratio of 0.0:2.0 and ratio of 2.0:0.0 as well as MS media supplemented with Kinetin:NAA with ratio of 0.0:2.0 and ratio of 0.5:1.0. This may occur due to experimental error, in which contamination may occur due to improper techniques while doing PTC such as lack of using aseptic techniques to make sure sterile condition during transferring the samples into the MS plate. Conclusion In conclusion, suitable medium supplemented with correct ratio of cytokinin and auxin are the most important key in order to get successful induction of organogenesis in vitro. Besides that, sterile environment is also one of the important factors in determining the successfulness of PTC and in order to obtain expected

Essay examples --

The Sibling Rivalry in Shakespeare's Taming of the Shrew Rivalry between family is an intense, emotional competition among siblings that can put them against one another to obtain approval, attention, or love. This is an aspect that has been frequently the plot of many plays, books and films. Taming of the Shrew, by William Shakespeare teaches the audience that preference within a family may led to potent hate when it comes to sisters. A play which embodies the rivalry between sisters Katharina and Bianca, produced by their fathers and spouse candidate preference toward Bianca. Shakespeare establishes a clear comparison between the sisters through their father, Baptista. From the beginning of the play we see a clear preference toward Bianca. He discusses how their daughter are to be in a relationship. â€Å"Gentlemen, importune me no farther,/That is, not to bestow my youngest daughter/Before I have a husband for the elder./If either of you both love Katharina,/ Because I know you well and love you well/ Leave shall you have to court her at your pleasure† (1.1.48-54). Baptista freely offers Katharina with out any detachment, while Bianca should be courted properly by the men. In addition to her father preferences, the mates wanting to marry the sisters also like Bianca. As one of the mates discussing which they prefer, Tranio refers to â€Å" [The] eldest sister is so curst and shrewd /That till the father rid his hands of her† (1.1.180). From the first scenes it is clear that there is rivalry between who is the better sister. Katharine was considered a shrew for the fact that she is strong willed and independent, very opposite to her younger sister, as she is described as caring and subservient. By the second act Katharina reacts to a ... ...shamed that women are so simple/ To offer war where they should kneel for peace/ Or seek for rule, supremacy and sway/ When they are bound to serve, love, and obey†(5.1.165-168). Katherina ends her speech by telling the women that their husbands are their lords, masters and keepers, that women owe their husbands the same loyalty a subject owes his king. The conflict between the sisters is an example of a tense relationships, physical violence and a continuous struggle for attention, affection and love of their father. Not until the end of the play we see Baptist being content with the results of his daughters new husbands. The relationship between Katharina and Bianca is a realistic in its portrayal of rivalry between siblings, and it allows the audience to connect with the theme of this play. The values of true sisterhood and how its can later effect their lives.

Productive and Counterproductive Behavior

Productive and Counterproductive Behaviors An organization will have employees with one of two behavior types. They can either be productive or counterproductive. The root cause of counterproductive behavior can be rooted in employee turnover, attendance issues, or lack of training. It is up to the organization to make an attempt to identify what the causes of their employees’ counterproductive behavior and attempt to find a workable solution. Productive Behavior and its Impact on an Organization Employers want productive behavior out of their employees because productive employees create revenue for the organization. However, employees tend to think just because they are doing â€Å"something† and not idle they are being productive. Their way of thinking would be incorrect. Productive behavior is â€Å"employee behavior that contributes positively to the goals and objectives of the organization† (Jex & Britt, 2008). As we can see merely doing something to occupy ones time is not being productive. Whether or not an employee is being productive can be measured by the cost associated with the employee’s performance and how effective they are. For example, there are two employees in a shoe store with similar sales figures and they sell the same number of shoes each month. The only difference in these two employees is that employee A works ten hours per day and employee B works a six-hour day. Employee B is the more productive employee because employee B accomplishes the same goal in fewer hours. There is a lower cost associated with employee B making him the more productive employee. Efficient employees are productive because they complete large amount of work in a short period of time. When employees are being productive they are concentrating on performing â€Å"core tasks† (Jex & Britt, 2008) and are bringing in revenue for the organization. An organization with productive employees will see less employee absences and in their turnover rate. Counterproductive Behavior Counterproductive behavior is â€Å"behavior that explicitly runs counter to the goals of an organization† (Jex & Britt, 2008). Employees who are counterprodtive may be employees who have issues with absenteeism or frequently tardy, substance abuse, they may steal from the employer, or simply have â€Å"ineffective job performance† (Jex & Britt, 2008). Any behavior that prevents an employee from conducting their job and producing for the employer is considered counterproductive. An Employee who take all day to complete a task because they are there all day and getting paid by the hour is an employee that is counterproductive. Another behavior that is counterproductive is an employee that uses unsafe practices at work. By not following safety rules employees can cause an injury to themselves or to another causing the employee to miss work. As previously stated, absenteeism is a counterproductive behavior. Strategies to Increase Productive Behavior To increase productive behavior in an organization and decrease counterproductive behavior an organization must investigate the cause. Sometimes the reason an employee is counterproductive are out of the employees’ control. For situations that are within the employees’ control, the organization must clearly outline their expectations for their employees. An organization must also outline the consequences for such negative behavior. Drug and alcohol testing and an attendance guidelines policy can be established. These policies will let the employee know what is expected of them and the consequences for not complying. The organization should also make efforts to understand why their employees are not coming to work (Associated Content, 2007). There are times when a personal issue can have its affect on an employees’ behavior. An organization can begin an employee assistance program that would offer counseling to employees if such a program were within the organizations budget. Lack of training and the appropriate tools for performing necessary job functions are factors that are out of the employees’ control. The organization needs to ensure all of their employees have all the proper tools. They can easily do a visual survey and question employees asking them what tools they could use to better perform their job function. Organizations must also ensure all of their employees have the proper training. If employees are not adequately trained they do not know how to perform their job functions. Organizations must adequately train or retrain if necessary any and all employees that require training. At times performing follow up training with all employees ensures employees are all on the same page as far as what their specific job functions are. Performance appraisals also need to be performed on all employees in order for them to know how they are performing in according with their job expectations. These performance appraisals will also serve as a tool to help the employee improve their performance and productivity. A performance appraisal should also be used to praise employees and let them know what they are doing that makes a positive impact on the organization (hrVillage, n. d. ). Both of productive and counterproductive behaviors have their impact on an organization. There are a number of reasons an employee can be counterproductive. Once the root cause has been identified an organization can begin to turn the counterproductive behavior into productive behavior. References Associated Content. (2007). Call Centers: 6 Steps to Improving Employee Attendance and Performance. Retrieved, March 29, 2010, from http://www. associatedcontent. com/article/189770/call_centers_6_steps_to_improving_employee. html? cat=31 hrVillage. (n. d. ). Employee Productivity. Retrieved, March 29, 2010, from http://www. hrvillage. com/human-resources/employee-productivity. htm Jex, S. M. &Britt, T. W. (2008). Organizational psychology: A scientist-practitioner approach, 2nd ed. New Jersey: John Wiley and sons.

Pornography Should Not Be Distributed Out The World

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