Course: Experimental Methods in Studies of Plant Defense Responses

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Course title Experimental Methods in Studies of Plant Defense Responses
Course code KBC/EMOR
Organizational form of instruction Exercise
Level of course Master
Year of study not specified
Semester Winter
Number of ECTS credits 7
Language of instruction Czech, English
Status of course Compulsory-optional
Form of instruction Face-to-face
Work placements This is not an internship
Recommended optional programme components None
Lecturer(s)
  • Jedelská Tereza, Mgr. Ph.D.
  • Dostálková Silvie, Mgr. Ph.D.
  • Janků Martina, Mgr. Ph.D.
  • Jahnová Jana, Mgr.
  • Sekaninová Jana, Mgr. Ph.D.
Course content
The exercises occur as block classes at the end of the winter semester. The subject guarantor informs the students about the progress of the exercises at the beginning of the semester. Exercise dates are sent to students at least 3 weeks before the start of block classes. Individual exercise tasks are focused on the processing of control and stressed plant samples and subsequent analysis by spectrophotometric, fluorimetric, electrophoretic, histochemical, and molecular biological methods: 1. Processing of frozen plant material (control plants and plants stressed by abiotic stress factors). Homogenization, extraction. 2. Work with plant cells. Determination of cell viability and cell death under stress factors (fluorescence microscopy, spectrophotometry). 3. Localization of reactive oxygen and nitrogen species in plant tissues (fluorescence microscopy). 4. Measurement of the antioxidant enzymes activity and antioxidant capacity (multifunctional reader). 5. Detection of heat shock protein in control and stressed plants (SDS PAGE, Western blotting and immunodetection). 6. Analysis of differences between control and stressed plants using molecular biological methods (PCR, qPCR).

Learning activities and teaching methods
Dialogic Lecture (Discussion, Dialog, Brainstorming), Observation, Demonstration, Training in job and motor Skils, Laboratory Work
  • Homework for Teaching - 100 hours per semester
  • Attendace - 104 hours per semester
Learning outcomes
Students will be introduced to and practically practiced with common experimental methods to study plant responses to stress conditions.
Practice and apply advanced techniques and methods of plant stress biochemistry and physiology and to utilize them creatively in practice.
Prerequisites
Students are expected to have the knowledge and skills obtained from successful completion of the course KBC/BCH Fundamentals of Biochemistry.

Assessment methods and criteria
Student performance, Systematic Observation of Student

1) 100% participation in practice (replacement of 2 absences for serious reasons, exceptional situations will be dealt with individually, e.g. hospital stay). Any absence must always be excused in writing (email) in advance before the start of the exercise. 2) Submitted and acknowledged protocol. Pairs of students working together on a given problem will submit one common report in electronic form. The completed protocol must be submitted no later than 10 days after the last exercise! Students will be awarded 30 points for a log submitted by the due date that is correctly completed. In case of failure to submit the protocol by the deadline, 15 points will be deducted. The protocol is corrected and graded after submission. The protocol is not subject to further corrections thereafter. 3) A written test focused on understanding the studied issue and the methods used. During the exercise, the student's readiness will be tested in the form of a written examination (5 questions from the theoretical and practical part of the task). Each correctly answered question is awarded 2 points. The maximum number of points is 10. If the student obtains 3 points or less in the test, he/she will be expelled from the exercise and will have to repeat the exercise on an alternative date. In case of obtaining 4-5 points the supervisor is obliged to examine the student orally, in case of ignorance of the given topic the student is banned from the exercise and has to repeat the exercise in an alternative term. Due to unpreparedness it is possible to replace the exercise only once. 4) The student must complete all requirements to pass the course and receive the required number of points for preparedness and submitted report. Once the protocol is accepted, the points earned and the requirements for credit for this course are evaluated, i.e. 100% attendance and a submitted and accepted protocol. Criteria for scoring: 75 - 100% points - Credit is awarded (1st regular attempt). 50 - 75% points - Student will write a credit test (test must be written for more than 70% of the points to be awarded credit, credit test can only be repeated once (2nd and 3rd regular attempts). < 50% points - Student will not be awarded credit, must re-enroll in the course the following year.
Recommended literature
  • Kesawat MS et al. Regulation of Reactive Oxygen Species during Salt Stress in Plants and Their Crosstalk with Other Signaling Molecules?Current Perspectives and Future Directions. 2023.
  • Mondal S et al. Crucial plant processes under heat stress and tolerance through heat shock proteins. 2023.


Study plans that include the course
Faculty Study plan (Version) Category of Branch/Specialization Recommended year of study Recommended semester
Faculty: Faculty of Science Study plan (Version): Biochemistry (2022) Category: Chemistry courses 2 Recommended year of study:2, Recommended semester: Winter