المبحث الخامس: زواجها
بعدما انتھت نازک الملائکۃ دراستھا في أمریکا عادت إلی بغداد وعینت أستاذۃ مساعدۃ في جامعتھا۔ فأصدرت عدۃ دواوین شعریۃ ونقدیۃ، وبعدہ فترۃ انتقلت من جامعۃ بغداد إلی جامعۃ البصرۃ، ثم تزوجت بالدکتور ’’عبدالھادي محبوبۃ في سنۃ 1962م۔ وزوجھا ھذا کان زمیلھا في جامعۃ بغداد في قسم اللغۃ العربیۃ في کلیۃ التربیۃ وکان عالماً لغویاً وأستاذاً سابقاً بجامعۃ الکویت، تزوجتہ نازک وھي تقرب الاربعین سنۃ من عمرھا۔
وعندما توفي زوجھا كانت ھي أیضاً مریضۃ وکانت مقیمۃ في القاھرۃ ولکنھا لا تعلم بوفاتہ المفاجئة، لأنھا کانت مریضۃ۔ لذلک لم یخبرھا أحد ولم یشأ أحداً نقل (النعي) إلیھا خوفًا من أن یضر بصحتھا، وبعد فترۃ من وفاتہ أخبروھا بأنّہ سافر إلی بغداد، وھي لا تعلم بأنھا الآن وحیدۃ وقد فارقھا رفیق حیاتھا، ویقول إبنھا البراق بأننا أخبرناھا بعد أسابیع من وفاتہ ودفنہ في القاھرۃ ورثتہ في قصیدۃ عنوانھا ’’أنا وحدي‘‘، ویبدو ھذہ آخر قصیدۃ قدمتھا نازک وتم نشرھا قبل وفاتھا۔
Mankind is scattered across the surface of the earth in the forms of different religious creeds, tribal affiliations, race, colour and nationalities. Yet they share some traits that are similar and universal to all human societies. These traits include love for children and respect for the elders. However, changing times have eroded some of these positive traits. The condition of senior citizens is not enviable in the West and those following in their footsteps. Children are not fulfilling their natural duty to their old parents, and quite often the elderly are forcefully kicked out of their own houses and accommodated in designated places for the old, run by the state welfare organizations.
Chilli veinal mottle virus (ChiVMV, genus Potyvirus) is a common, wide spread, destructive and economically important plant virus that cause systemic infection in chilli resulting in mottling, vein clearing and leaf distruction. The successful production of chilli crop in Pakistan is mainly hampered due to ChiVMV which resulted in quite low average yield as compared to other countries. In Pakistan, limited work is only confined to occurrence and prevalence of ChiVMV from capsicum crop but no systemic and molecular study has been conducted on this virus. Development of control strategies and/or eliminate of viral diseases in plants requires better understanding and knowledge on the molecular characterization including sequencing and genetic diversity. Pathogen derived resistance (PDR) is one of the important strategies to implement effective and sustainable control measures against major viral diseases. The present research work was conducted to elucidate the genetic variability of Pakistani ChiVMV isolate and study the role of CP gene in conferring the resistance against Pakistani isolate of ChiVMV in model plant, Nicotiana benthamiana. In this regard Nicotiana benthamiana plants were transformed by using Agro-infiltration and stable transformation methods. Infectivity assay was done to evaluate the resistance level in transformed plants. Local lesion assay and host range studies were performed for biological purification, propogation and to evaluate the response of Pakistani ChiVMV isolate to different hosts. The 864 bases of CP gene was amplified in RT-PCR. CP cDNA was successfully cloned in pTZ57R/T TA cloning vector (p01). CP gene specific primers was designed with BglII and BsteII restriction sites. Using the clone p01 as template, 1kb fragment of CP gene was cloned with restriction sites (clone p02). CP cDNA from p02 was inserted in pcambia1301 after digestion with BglII and BsteII in sense or anti-sense direction under the control of CaMV35S promoter and transformed into Agrobacterium strain LBA4404. This transformed Agrobacterium culture was used for tansforming the Nicotiana benthamiana plants through agro-infiltration and by stable transformation methods. Confirmation of the transformation was carried out through PCR amplification of CP and hygromycin gene. Resistance level in transformed Nicotiana benthamiana plants against ChiVMV isolate ATIPK was confirmed through infectivity assay. Local lesions were produced by Pakistani ChiVMV isolate in Chenopodium quanoa plant. Different host plants like chilli varities, Datura metal and Nicotiana tabaccum showed the same symptoms for ChiVMV isolate ATIPK as reported for other ChiVMV isolates from the different areas of the world. A total of 986 nucleotides were obtained from each clone consisting of 96 basis of 3’ end of Nib gene, full length CP gene (864) and 23 bases from the 3’UTR. The sequence of ChiVMV(ATIPK) isolate was deposited to Genebank (acc. No. KJ472764). A pairwise comparison of ATIPK CP sequence with 22 highly matched sequences from database revealved that it has the highest nucleotide identity with an isolate from Thailand DQ854956 i.e 88.4% and lowest nucleotide identity was 86.8% with an isolate from India (DQ854964). Whereas highest amino acid identity was 91.8% with 3 isolates from India (EF213679, EF213681 and EF213703) and lowest amino acid identity was 89% with an isolate from Indonasia (AB703256). Putative transgenic N. benthamiana plants using CP gene of Pakistani ChiVMV isolate ATIPK, were used for the evaluation of resistance level against same isolate. Inoculated 2nd generation plants (T1-T8) indicated the presence of CP gene by overcome the infection and further confirmed through DAS ELISA test.The ELISA readings confirmed the phenotypic symptoms and indicated the presence of less amount of titer in transgenic Nicotiana benthamiana plants as compare to control plants. These findings on characterization and expression of understudied gene may enable us to define the position and role of it in the resistance that would be useful for developing effective control measures against this menaces threat (ChiVMV) to the solanacious crops.