Keragaman Genetik Dan Pendugaan Jumlah Gen Ketahanan Kacang Panjang (Vigna Sinensis L.) Terhadap Penyakit Kuning

Penyakit kuning pada kacang panjang berdampak pada penurunan produksi. Gejala serangan diawali dari gejala daun keriting serta mengakibatkan polong berwarna kuning. Penelitian ini bertujuan mengetahui nilai heritabilitas dan ragam genetik serta menduga jumlah gen pengendali ketahanan kacang panjang terhadap penyakit kuning. Penelitian dilaksanakan di Kabupaten Kediri pada bulan April sampai Juli 2013. Bahan penelitian adalah populasi UB 715 A (P1), Hitam Putih (P2), populasi F1 dan populasi F2. Berdasarkan hasil penelitian, populasi UB 715 A (P1 ) menunjukkan respon tahan terhadap penyakit kuning, populasi Hitam Putih (P2) menunjukkan respon rentan, dan populasi F1 dan F2 menunjukkan respon sedang. Karakter jumlah polong dan jumlah biji per tanaman memiliki keragaman yang sempit sedangkan karakter panjang polong, bobot segar polong, umur berbunga, dan umur panen memiliki keragaman yang luas. Karakter panjang polong dan jumlah biji per polong memiliki nilai heritabilitas rendah, sedangkan karakter jumlah polong, bobot segar polong, umur berbunga, dan umur panen memiliki nilai heritabilitas tinggi. Rasio sifat ketahanan terhadap penyakit kuning pada populasi F2 adalah 9 tahan : 3 sedang : 4 rentan yang berarti ketahanan terhadap penyakit kuning dikendalikan oleh dua gen dengan aksi gen epistasis resesif

Mid-term outcomes of biventricular obstruction and left ventricular outflow tract obstruction after surgery correction in child and adolescent patients with hypertrophic cardiomyopathy - Fig 2

<p>Fig 2a. Preoperative two-dimensional transthoracic echocardiography (tte) parasternal long axis (PLAX) views in a 16-year-old hypertrophic cardiomyopathy patient with BVOTO. (A) PLAX view demonstrating the massive septal hypertrophy and the thickening of the ventricular septum bulging into the LVOT and RVOT resulting in biventricular obstructions (the colour flows). (B) Colour Doppler flow imaging of PLAX view during systole showing high velocity jet flow simultaneously in both LVOT and RVOT. Postoperative PLAX views showing a substantial decrease in the ventricular septum thickness and an increase in the RV and LV cavity sizes during diastole (C) and the LV and RV colour flows showing laminar without evidence of significant residual obstructions during systole (D).RV: right ventricle; RVOT: right ventricular outflow tract; IVS: interventricular septum; LV: left ventricle; LA: left atrium; LVOT: left ventricular outflow tract.AO: aorta. Fig 2b. Preoperational cardiovascular magnetic resonance (CMR) image 3-chamber views during diastole (A) and systole (B) showing remarkable myocardial hypertrophy at the base ventricular level with LVOT and RVOT obstruction. The postoperative CMR images (C, D) showing thinner IVS, wider LVOT and RVOT diameter and larger LV and RV cavity without the projection of septum into RVOT or LVOT after biventricular resection. LA: left atrial; LV: left ventricular.</p

Baseline characteristics.

<p>Baseline characteristics.</p

Middle-Term results in the three groups.

<p>Middle-Term results in the three groups.</p

Single Channel Recordings Reveal Differential β2 Subunit Modulations Between Mammalian and <i>Drosophila</i> BK_Ca(β2) Channels

<div><p>Large-conductance Ca²⁺- and voltage-activated potassium (BK) channels are widely expressed in tissues. As a voltage and calcium sensor, BK channels play significant roles in regulating the action potential frequency, neurotransmitter release, and smooth muscle contraction. After associating with the auxiliary β2 subunit, mammalian BK(β2) channels (mouse or human Slo1/β2) exhibit enhanced activation and complete inactivation. However, how the β2 subunit modulates the <i>Drosophila</i> Slo1 channel remains elusive. In this study, by comparing the different functional effects on heterogeneous BK(β2) channel, we found that <i>Drosophila</i> Slo1/β2 channel exhibits “paralyzed”-like and incomplete inactivation as well as slow activation. Further, we determined three different modulations between mammalian and <i>Drosophila</i> BK(β2) channels: 1) dSlo1/β2 doesn’t have complete inactivation. 2) β2(K33,R34,K35) delays the dSlo1/Δ3-β2 channel activation. 3) dSlo1/β2 channel has enhanced pre-inactivation than mSlo1/β2 channel. The results in our study provide insights into the different modulations of β2 subunit between mammalian and <i>Drosophila</i> Slo1/β2 channels and structural basis underlie the activation and pre-inactivation of other BK(β) complexes.</p></div

Single channel recordings revealed the enhanced pre-inactivation in dSlo1/β2 channel.

<p><b>(A)</b> Representative single channel recordings of mSlo1/β2 from inside-out patches in the presence of 10 μM Ca²⁺ using a depolarizing voltage of 100 mV as indicated. The scale bars were 100 ms and 25 pA, respectively. <b>(B-C)</b> Three representative single-channel traces of mSlo1/β2(W4E) and mSlo1/β2(W4E,D16R,E17K) recorded from inside-out patches at +100 mV in 10 μM Ca²⁺. Bottom traces showed ensemble averages of 100 consecutive sweeps. Initial capacitive currents within the first 0.5 ms of voltage steps were deleted. Right, histogram of open-time durations from the two single-channel recordings. <b>(D-E)</b> Three representative single-channel traces of dSlo1/β2 and dSlo1/β2(D16R,E17K) recorded from inside-out patches at +100 mV in 10 μM Ca²⁺. Bottom traces showed ensemble averages of 100 consecutive sweeps. Right, histogram of open-time durations from the two single-channel recordings. <b>(F)</b> Immunofluorescence imaging of dSlo1-GFP/β2(D16R,E17K)-72Myc at different α:β transfection ratios of 1:4.</p

β2(K33,R34,K35) caused slow activation in dSlo1/Δ3-β2 channel.

<p><b>(A)</b> Top panel was the construct for β2 subunit truncations Δ3-β2, Δ30-β2, and Δ35-β2. Bottom panel showed the traces recorded at 100 mV in 10 μM Ca²⁺ solution for dSlo1 (black), dSlo1/Δ3-β2 (orange), dSlo1/Δ30-β2 (dark green), dSlo1/Δ35-β2 (pink), and dSlo1/Δ30-β2(K33,R34,K35) (light blue). <b>(B)</b> Average activation time constants (τ_a) for dSlo1 (68.55 ± 6.65 ms), dSlo1/Δ3-β2 (288.05 ± 74.01 ms), dSlo1/Δ30-β2 (623.37 ± 74.18 ms), dSlo1/Δ35-β2 (92.48 ± 25.88 ms), and dSlo1/Δ30-β2(K33,R34,K35) (127.92 ± 26.22 ms). Traces were fitted with a single exponential equation. <b>(C–D)</b> Single channel recordings and current histograms for the dSlo1/Δ3-β2 and dSlo1/Δ30-β2 channels. The currents between the blue lines were the raw data used for histogram analysis and the histogram were fitted with triple or double Gaussian equations.</p

Cartoon schematic to clarify the different modulations of β2 subunit between mammalian and <i>Drosophila</i> BK_Ca(β2) channels.

<p>Gray and orange bars indicated dSlo1 or mSlo1 and β2 subunit, respectively. Blue, orange, and pink circles indicated the α and β2 binding or interactions, β2(K33,R34,K35), and β2(D16,E17) or pre-inactivation, respectively. The black circle indicated the enhanced pre-inactivation in dSlo1/β2 channel. Firstly, in dSlo1/β2 channel, the inactivation ball FIW is not enough for the complete inactivation, and may suppress the α and β2 binding or interactions. Secondly, β2(K33,R34,K35) delays the pore opening (or voltage sensor movement). Thirdly, dSlo1/β2 channel has enhanced pre-inactivation than mSlo1/β2 channel.</p

dSlo1/β2 channel doesn’t have complete inactivation.

<p><b>(A)</b> Macroscopic currents of mSlo1, mSlo1/β2, mSlo1/Δ3-β2, dSlo1, dSlo1/β2, and dSlo1/Δ3-β2 obtained from inside-out patches in the presence of 10 μM Ca²⁺ according to the protocol indicated. Scale bars represent 50 ms and 2 nA for mSlo1/x channels, and 200 ms and 0.5 nA for dSlo1/x channels. The α:β transfection ratio was 1:8 for dSlo1/ β2, and 1:0.2 for other combinations. The voltage ranges of the test potential were from -160 mV to +160 mV for mSlo1, mSlo1/β2, and mSlo1/Δ3-β2, and from -160 mV to 140 mV for dSlo1, dSlo1/β2 and dSlo1/Δ3-β2. <b>(B)</b> β2 binding percentages of a series of α/β2 channels at different α:β transfection ratios. Gray bars were obtained from patch clamp recordings and the red bars from immunofluorescence imaging. The mSlo1/β2 channel currents were analyzed based on patches with entire inactivation, mSlo1/Δ3-β2 based on the G-V relationship left shift, dSlo1/β2 based on inactivation or briefly opening recordings, and dSlo1/Δ3-β2 based on slow activation. dSlo1-GFP/β2-72Myc at [1:1] (N = 60) and at [1:4] (N = 61) were analyzed based on surface immunofluorescence images of β2-72Myc and then normalized against the dSlo1/β2 [1:4] results obtained from patch clamp recordings. The error bars represent the standard error of the mean (SEM). <b>(C)</b> Immunofluorescence imaging of dSlo1-GFP/β2-72Myc at different α:β transfection ratios of 1:1 (left) and 1:4 (right). Scale bar represents 10 μm.</p

Single channel recordings of mSlo1 and dSlo1 combinations with β2 subunit.

<p><b>(A)</b> Representative single channel recordings of mSlo1, mSlo1/β2, and dSlo1 channels from inside-out patches in the presence of 10 μM Ca²⁺ using a depolarizing voltage of 100 mV, as indicated in (B). The scale bars were 100 ms and 25 pA, respectively. <b>(B)</b> Representative single channel recordings of the dSlo1/β2 channel before (top) and after (bottom) applying 0.2 mg/ml trypsin. The bottom black traces are ensemble averages from 100 consecutive sweeps and its enlarged trace. Red traces are single exponential fits to the black traces. The inactivation τ_i and activation time τ_a constants were 166.2 ms and 120.8 ms, respectively. The α:β transfection ratio was 1:8. The scale bars were shown as indicated. <b>(C)</b> Representative single channel recordings of the dSlo1/Δ3-β2 channel before (top) and after (bottom) applying 0.2 mg/ml trypsin. The bottom black traces are ensemble averages from 100 consecutive sweeps. Red traces are single exponential fits to the black traces. The activation time constants τ_a were 1446.3 ms and 168.1 ms, respectively. The α:β transfection ratio was 1:8. The scale bars were 100 ms and 25 pA as shown in (B).</p